ALBERT

Notes: A healthy 22-year-old woman was noted to have erythrocytes of the Pk phenotype: a strong Pk antigen, no detectable P antigen and anti-P antibody in her serum. Her erythrocytes contained four to six times as much Pk glycolipid (globotriaosylceramide or CTH) and approximately half as much P glycolipid (globotertraosylceramide or globoside) as normal red cells. The structures of CTH and globoside were characterized by analysis of permethylated sugars and complement fixation in addition to chromaographic mobility and sugar composition. Inasmuch as the erythrocytes of two Pk individuals that were analysed previously (Marcus et al., 1976) contained no detectable globoside, these abnormalities appear o represent a new phenotype in the P blood group system.

Notes: We studied 201 unrelated French Basque individuals for HLA and Bf polymorphisms. The haplotypes of eighty-seven of them were deduced from family studies. The results show the frequency of the Bf F1 allele (0.1393) which is the highest one currently reported. They confirm the high frequencies of HLA-Aw19.2 and B18 previously reported in that population and show that a whole haplotype with strong linkage disequilibria, namely Aw19.2, Cw5, B18, Bf F1, DRw3 is frequent. On the other hand, the gene frequency of Bf S is decreased (0.5497) as compared with the other European Caucasoïd populations, while a slight increase in the Bf F gene frequency (0.2960) appears. These results point out that it is of importance to consider the genetic background in choosing the population where linkage disequilibria are to be studied.

Notes: The genetic control of hybrid resistance to BALB/c fibrosarcoma Meth-A was investigated. A Meth-A tumour grew slower in (BALB/c X C57BL/6)F1 and reciprocal hybrid mice than in syngeneic BALB/c mice and was also found to grow slower in females than in males. Significant F1 resistance was demonstrated after both subcutaneous and intraperitoneal injection of tumour cells. However, (BALB/c X DBA/2)F1 mice did not show any significant resistance to Meth-A. In H-2 linkage studies of [BALB/c X (BALB/c X C57BL/6)] backcross mice, no statistically significant differences in the resistance of H-2 heterozygotes and homozygotes to Meth-A were observed. These results indicated that F1 hybrid resistance to Meth-A was controlled by non-H-2-linked resistance factor(s). No linkage was observed between resistance to Meth-A and coat colour c- and b-loci.

Notes: CBA/N mice have an X-linked B cell defect which prevents them from responding to non-mitogenic thymic independent (TI-II) antigens such as dinitrophenylated (DNP-AGG) Ficoll. The F1 male progeny of CBA/N female mice express the same defect. Spleen cell suspensions from such defective mice (CBA/N X C3H/HeN F1 males) could not respond to DNP-AGG-Ficoll following in vitro immunization and subsequent transfer into irradiated, syngeneic, F1 male recipients as expected. In contrast, normal CBA/N X C3H/HeN F1 female spleen cells could respond and effect a ‘rescue'; they mounted strong plaque-foriming cell 7 days after in vitro exposure to DNP-AGG-Ficoll and subsequent transfer into irradiated F1 male recipients. Defective F1 male spleen cells could bind significant quantities of DNP-AGG-Ficoll, however, after, in vitro exposure. Extensive washing of these spleen cells could not reverse this binding. Such DNP-AGG-Ficoll-exposed and washed F1 male spleen cells could, after transfer, aid normal untreated F1 female cells in their rescue function. The defective F1 male spleen cells could convey immunogenic quantities of DNP-AGG-Ficoll to the ‘rescuing’ F1 female cells.Mitomycin treatment of F1 male cells did not interfere with their conveyor function. Goat anti-mouse μ serum impeded the passive antigen conveyor function of defective F1 male cells as did prior exposure to high concentrations of free DNP-AGG hapten. Our data support the view that the B cell defect of CBA/N X C3H/HeN F1 male mice does not relate to antigen binding, but rather to an inability to be effectively triggered by certain cell-bound polymeric antigens.

Notes: Results of a population study with all currently available B-cell specific alloantisera indicate that eight antigens controlled by the RhLA-linked DR locus can now be identified. This leaves a gene frequency of about 0.15 for unidentified or ‘blank’ antigens of that locus. Of the nine identifiable la antigens which are not controlled by the DR locus, three or four may form the basis of a second series which is probably also controlled by the RhLA region.

Notes: Induction of tolerance to bovine serum albumin was studied in mice selected for high (H) or low (L) antibody responsiveness and in their F1 hybrids. No high or low zone tolerances were obtained in H mice whereas L mice were susceptible to tolerance induction by the two schedules. H mice were immunized by repeated injections of tolerogenic BSA for low zone tolerance induction but not after the administration of a single high dose of tolerogenic BSA. Resistance to tolerance induction is dominant in F1 hybrids.

Notes: An attempt is made to account for immunoglobulin chain synthesis in terms of genetic events involving IS or controlling elements analogous to those found in bacteria, maize and drosophila. Transposition of variable and constant genes and normal immunoglobulin chain synthesis as well as qualitative and quantitative abnormalities might be explained by such regulatory elements. Intrachromosomal transpositions over short distances would be expressed as apparent hypermutability or redundancy of the variable DNA segment. The constant gene might comprise four sequences coding for the three homology domains and the hinge, separated by intervening sequences. A strong preference for shortrange transposition on the same chromosome and immobilization of the controlling element in the end might account for allelic exclusion.

Notes: Fab2 fragments from antisera raised in rabbits with partially purified cellular and serum HLA antigens were tested for their ability to block the cytolytic activity of operationally specific HLA-A, B alloantisera. One Fab2 fragment preparation blocked the cytolytic activity of all the HLA-A,B alloantisera tested; the remaining nine inhibited the lytic activity of alloantisera to certain HLA-A,B allospecificities, suggesting that these xenoantisera contain antibody to certain HLA-A,B allotype determinants or to closely associated structures. In contrast to previous reports in the literature none of the xenoantisera contained significant amounts of antibodies to human β2-microglobulin.

Notes: NIM-M8 is a monoclonla IgM antibody, specific for the LWab antigen as shown by its reaction with red cells of all donors except those lacking LWa, LWb and LWab. Indirect immunofluorescent staining and cell sorter analyses have shown that LWab is present on a subpopulation of human lymphoctes. Cell fractionation studies indicate that subsets of both B and T cells express LWab and it may, therefore, provide a further marker for heterogeneity in these lymphocyte populations.

Notes: Immunoprecipitation using a monoclonal antibody showed that the Wrb antigen is present on the abnormal (δ-α) hybrid sialoglycoprotein of Sta-positive human erythrocytes but not on the abnormal (δ-α) hybrid sialoglycoprotein of Dantu-positive erythrocytes. These results provide further information regarding the nature and location of the Wrb antigen on the normal erythrocyte sialoglycoprotein α.

Notes: We studied structural and functional characteristics of lymphocytes from adult and fetal baboons (Papio cynocephalus). Flow cytometry with monoclonal antibodies to human lymphocyte antigens and plant lectins was used to define expression of surface antigens on lymphocytes from adult and 140 day fetal baboons (term = 180 days). Major T cell antigenic determinants on adult and fetal baboon lymphocytes were the Tp50, Tp32-45, and p45 glycoproteins detected by monoclonal reagents T11, OKT8, and OKT10 respectively. Baboon T lymphocytes did not react with the OKT3/anti-Leu4 or OKT4/ anti-Leu3a reagents which detect, respectively, Tp19-29 and Tp55, major surface glycoproteins on human T lymphocytes. OKT6, which identifies the human TL antigen equivalent on thymocytes, did not react with baboon thymocytes. These data demonstrate major evolutionary divergence between human and baboon T lymphocytes. By contrast, baboon lymphocytes resembled human peripheral lymphocytes in reactivities with several non-T cell reagents. Lectin binding studies revealed substantially fewer peanut agglutinin-and wheat germ agglutinin-binding cells in suspensions of baboon fetal splenocytes and adult peripheral lymphocytes compared with fetal thymocytes. Thereffore, maturation of baboon T lymphocytes is associated with loss of surface carbohydrate structures that bind these lectins. Adult and fetal baboon lymphocytes resembled human and murine lymphocytes in their capabilities to respond to mitogens and to produce interleukin-2. As in oter species, adult, but not fetal baboon lymphocytes, mediated NK activity against a variety of nucleated target cells. Despite divergence in lymphocyte antigen epression, babbon lymphocyte functional development colsely parallels that seen in humans.

Notes: The Gm, Am and Km immunoglobulin allotypes and ABO blood groups were studied in three groups of Tunisian Berbers.The results showed that the actual Berbers of Tunisia present certain heterogeneity and their ancestors were probably the first inhabitants of North Africa. Indeed, although their Gm-Am haplotypes are mainly Caucasoid, some of them are typically African.The group of Kesra village, the most Caucasoid, shows frequencies of Gm-Am haplotypes very close to those of South European populations, particularly the Spanish, who are probably of the same origin. The gene frequencies of the ABO groups in the three Berber groups were similar to those recorded in European populations with a relatively high frequency of the O genes typical of the Berbers.

Notes: Monoclonal antibody 212.i.4.2 mediated complement-dependent lysis of spleen and lymph node cells carrying the tw1, tw12, tw71, t6, tw73, and tLub1 haplotypes, while cells from mice carrying 11 other t haplotypes were not lysed. The antibody also detected an epitope controlled by genes in the H-2Dd region of non-t mice. A molecule of 46,000 molecular weight was immunoprecipitated by 212.i.4.2 from detergent extracts of 125I-labelled spleen cells of +/tw12 and B10.D2 mice. The H-2dm2 mutation did not alter the expression of the epitope recognized by 212.i.4.2. However, the H-2dm1 mutation decreased the reactivity of lymphoid cells with the antibody in cytotoxicity tests, and 212.i.4.2 immunoprecipitated little or no protein from extracts of B10.D2(R106) spleen cells which carry the H-2dm1 mutation.

Notes: Spleen cells from Balb/c mice given multiple injections of intact human erythrocytes (group O, NN) were fused with NS1 myeloma cells. Culture fluids from the resulting hybrid cells were screened for agglutinating antibody against a panel of erythrocytes. One cell line, 2/23, secreted an IgM antibody which reacted more strongly with NN than with MM cells. Neuraminidase or papain treatment of erythrocytes abolished agglutination whereas trypsin treatment did not. Reactions with U-erythrocytes of different MN phenotypes confirmed the anti-N specificity of monoclonal antibody 2/23. This is the first report of monoclonal anti-N stimulated by the immunization of mice with intact erythrocytes.

Notes: Spleen cells from 30 individual murine irradiation chimeras of the type (P1 x P2)F1→ P1 were compared in a rosetting assay for H-2K and H-2D cell surface antigen expression with normal (P1 x P2)F1 hybrid controls. Eleven out of the 30 chimeras were in the normal range, but the other 19 differed from F1 controls by 4- to 100-fold in endpoint titre for at least one H-2K or H-2D antigen. Every possible class of variation was found, i.e. up or down variation of H-2K or H-2D antigens of P1 or P2 type. This evidence, together with data from T6 chromosome marker experiments which also showed full reconstitution of lethally irradiated P1 recipients by (P1 x P2)F1 donor lymphomyloid stem cells, suggested that incomplete reconstitution was not the cause of H-2 antigenic variation.Low expression of P2 H-2 antigens on spleen cells derived from (P1 x P2)F1→ P1 chimeras was investigated further. Fifteen lethally irradiated (P1 x P2)F1 recipients of bone marrow cells from two such chimeras were all of normal F1 H-2 phenotype when tested 10-12 weeks after reconstitution, thus excluding stable, low P2 H-2-expressing variant F1 stem cells as a cause of the phenomenon. If P1 recipients were hyperimmunized against P2 cells before lethal irradiation and reconstitution with (P1 x P2)F1 stem cells, there were significantly fewer Till-McCulloch colonies in their spleens 10 days after reconstitution than in spleens of unimmunized controls. Also 〉 90% of immunized recients died by 6 weeks after stem cell injection but two survivors both showed very low levels of P2 H-2K and H-2D antigens. These results together with previously published evidence of anti-P2 Tc cell activity and P2 skin graft rejection in (P1 x P2)F1→ P1 chimeras suggested that residual anti-P2 immunological capability in lethally irradiated P1 recipients may be associated with low P2 H-2 expression on their F1-derived spleen cells, although the mechanism does not involve selection of stable, variant F1 stem cells. The mechanism(s) of other classes of variation in H-2 expression in (P1 x P2)F1→ P1 chimeras were not investigated.

Notes: RNA was extracted from the splenocytes of Brucella abortus antigen stimulated mice and of control mice. The proportion of chromatographically separated polyadenylated 11.2S mRNA, was determined. With the technique used, only stimulated mice exhibited significant amounts of this RNA species. The highest level was reached 1 day after the stimulation, and the decay from this level presented an oscillatory form during the 4 weeks following the injection.In two different genetic backgrounds, H-2b mice did not respond to the stimulus, in contrast to H-2a and H-2f mice. H-2b/H-2f heterozygotes behaved roughly as intermediate between H-2b and H-2f mice. This genetic control seems to parallel the genetic control of some Brucella-induced, thymus-dependent events previously described.

Notes: Book Reviews in this ArticleR. WITKOWSKI and O. PROKOP: Genetik erblicher Syndrome und Mgbildungen. Worterbuch fur die Familienberatung.

Notes: Monoclonal anti-Ia inhibition experiments were conducted to confirm and extend genetic mapping data of I-A gene control of immunity to human haemoglobin (Hb). It was found that the Aβ gene is of critical importance in conferring immunity to the α-chain and β-chain subunits of Hb. A possible involvement of I-E region genes in B10.D2 mice to β-chain is discussed. Through the use of an α-chain specific T cell clone data, is obtained indicating that an intact Ia.8+ Aβ chain is necessary for antigen presentation in vitro.

Notes: Immunoglobulin was obtained from a hybridoma cell line, which was a reclone of a hybrid between a rabbit cell and mouse myeloma cell (X63-Ag8). The immunoglobulin, isolated from the cell culture medium was found to be homogeneous by isoelectrofocusing and immunoelectrophoresis and consisted of mouse heavy and rabbit light chains, linked by disulphide bonds. All immunoglobulin molecules carried both mouse and rabbit determinants; mouse determinants were associated only with the heavy chains while rabbit determinants were only associated with the light chains. The rabbit light chains were of Ab4 allotypical specificity, but possess only some of the Ab4 determinants normally present in Ab4/Ab4 animals. It was sugested that the restriction in allotypical specificity may be a general property of light chain Ab allotypes; the normal serum immunoglobulins may be heterogeneous with respect to the allotypic determinants and any one molecule may possess only a proportion of determinants detected by a conventional anti-allotype antiserum.

Notes: In a family study of coeliac disease, HLA types in fifty-three patients and their relatives were examined. There are no differences in HLA frequencies between child and adult patients. Comparison with a random series of normal controls shows increases frequencies in patients of HLA-A1 and B8, while the family material shows that there is also an excess of haplotype I-8. The excess of homozygotes is thought not to be a factor in the aetiology. Intrafamilial analysis shows that only B8 is significantly associated with the disorder. It is argued that the HLA association does not indicate a ‘coeliac gene’ but that the B8 allele is a major gene in a polygenic system affecting the disorder.

Notes: Several interesting and clinically important strains of Mus musculus were serotyped with alloantisera recognizing the different alleles of the Thy-1, Ly-1, Ly-2, Ly-3, Ly-4, Ly-5, Ly-6 and Ly-7 loci. The strains were the Biozzi high and low responder strains, of importance in immune responsiveness studies, and the strains BXSB, MRL and MRL—lpr/lpr—strains which spontaneously develop immune complex disease. In addition the related species Mus musculus castaneus was typed with the same reagents. The knowledge of the cell surface phenotype should prove useful in various functional studies involving these interesting strains.

Notes: Mouse strains differ widely in their natural killer(NK)-cell activity. In the (A X B6) X A backcross, high reactivity was linked to H-2b, although non-H-2-linked genes were also demonstrated (Petranyi et al., 1975). Harmon et al. (1977) demonstrated an H-2Dd-associated reactivity gene (1977). In the present study, we have tested eleven B10 congenic strains for NK activity. The H-2Dd strains B10.A, B10.T(6R), B10.S(7R), B10.HTT and B10.D2 were more highly reactive than B10, B10.S, B10.G, B10.A(2R) and B10.BR, which do not carry the d allele at the H-2D locus. While this confirms the H-2Dd association of a reactivity gene, an exception was found in the B10.A(5R) strain that was low reactive in spite of the fact that it carries H-2Dd. This suggests the possibility that the H-2Dd-associated gene is outside H-2, to the right of Tla.The AKR.H-2b congenic line had the same low activity as the AKR.H-2k strain; both were much lower than B6. This suggests either one of two possibilities: the H-2b-linked reactivity gene is relatively distant from the H-2 complex, although localized on chromosome 17 or alternatively, if localized within or in the close neighbourhood of H-2, it requires non-H-2 genes for full expression.Previously, we have shown that the B6 X DBA/2 F1 hybrid was more highly reactive than either one of its two parental strains (Klein et al., 1978). A similar complementation effect is described in the present paper for the B10.D2 congenic strain. The high reactivity of this line can be due to the combined effect of an H-2b-linked gene from DBA/2 and the non-H-2 background of B10 or, alternatively, the former, together with an H-2b-linked gene from B6 that lies well outside outside the H-2 locus itself.

Notes: The K region of the H-2 major histocompatibility complex (MHC) of mice of H-2Kk haplotype codes for two distinct alloantigens. One of these alloantigens, designated k-common, is expressed by C3HfB/HeN mice (C3Hf). The other alloantigen, designated k-unique, is not expressed by C3Hf mice. The H-2 haplotype of C3Hf mice has been classified as kv1 and the variant antigen distinguishing this strain from mice of H-2Kk haplotype has been designated kv1-unique. Several transplacentally-induced lung tumours of C3Hf mice express the k-unique, rather than the expected kv1-unique, antigen. The immunogenicity of the k-unique antigen on C3Hf-derived lung tumours varies with different tumours. In particular, the capacity of the k-unique antigen to induce radioresistant immunity in C3Hf mice appears to be lost on long term cultured tumour cells even though the tumour remains susceptible to in vivo immune responses directed against the k-unique antigen. Alterations in expression and in immunogenicity of unique H-2-coded antigens may dictate the nature and efficacy of immune surveillance of autochthanous tumours.

Notes: The inbred mouse strains BN/a and BN/b have haplotype H-2bp characterized by H-2.33 and lacking any other private specificity known in inbred strains. Two transplantable B cell leukaemias which originated in BN/a and BN/b mice treated with anti-lymphocyte globulin were tested serologicaly for H-2 antigens. Tests during passages 169-181 revealed several quantitatively different reactions with sera against public specificities, some of these being due to Ia antibodies. No change in expression of the private specificities was seen. On the other hand, during the later passages (239-258) a number of qualitative differences were seen, the most remarkbale being the loss of H-2.33 and gain of H-2.4, 31. The overall serological pattern of cells resembled that of H-2d rather than of H-2bp haplotyes and this was confirmed by absorption tests. The changes reported here may be due to alterations of repression and derepression pattern of the presumed multiple structural H-2 genes present in the genome.

Notes: Cellular cytotoxicity experiments were done to test the potential of the extra H-2 antigenic specificities on the K36 spontaneous leukaemia as targets. In addition, experiments were performed to rule out the possibility that the target determinants could be normal cross-reacting alloantigens, e.g., T1, Qa, non-H-2 and previously undetected H-2 public specificities, which are more readily detectable on tumours than on normal cells.We used F1 hybrid mice, in which one parent was of the strain of origin of the tumour (K36) and the other parent of the B10 congenic series, i.e., (AKR x B10)F1. These cells were stimulated by lymphoid cells from other B10 congenic strains, B10.A and B10.D2, and tested against the test tumour K36 and several PHA blast controls. Several K36 sublines as well as a cloned line of K36 (K36.16) were used and significant cytotoxicity against. an H-2d-like target on these tumour cells was obtained. These data exclude the possibility of a corss-reactive alloantigen, e.g., undetected H-2 public specificity, or differentiation antigens. These results with the K36 tunour were consistent with our immunochemical studies (see Schmidt & Festenstein, 1980) and were confirmed and extended by cold target inhibition experiments. In these experiments, B10.BR cells were sensitized by B10.D2 lymphoid cells and tested against B10.D2 (51chromium-labelled PHA balsts). Two kinds of normal unlabelled lymph node suspensions as well as the K36.16 tumour cell suspension were used. Significant specific inhibition of between 19% and 40% was obtained using K36 and between 23% and 37% using B10.D2 (positive conrol). AKR cells (negative control) in contrast were unable to reduce the precentage specific cytotoxicity.Since it was already known that the H-2Kk gene products are missing from this tumour (Schmidt & Festenstein, 1980), it was of interest to test whether cytotoxic effectors directed against the H-2kk gene products were able to kill the K36 tumour. Accordingly, B10.D2 lymphoid cells were sensitized to B10.BR (C3H.0H) and B10.A targets, respectively, and tested against K36 and appropriate controls. Only weak killing was observed when sensitization was effected against the K end of the H-2k haplotype (i.e., using B10.A as the sensitizing cell) but strong and significant cytolysis was found when the sensitization was against the whole H-2k haplotype or against the H-2Dk gene product. These results were confirmed by cold target inhibition studies.These experiments provide further indications for the H-2d-like characteristics of these allodeterminants. We have already excluded some of the possible explanations for these findings (i.e., corss-reactions with H-2 and non-H-2 normal specificities). The cold target inhibition experiments rule out non-specific viral effects. Thus, we favour an alteration in regulatory genes leading to repression of the H-2Kk product and derepression of the H-2Dd product, but cannot formally rule out highly corss-reactive H-2-like virally encoded determinants.

Notes: Alien H-2k-like antigens were found to be expressed by a methylcholanthrene induced tumour of BALB/c (H-2d) origin. H-2 specificities of the k haplotype were detected on this tumour by a variety of serological techiques, including 51Cr-release cytotoxicity, microradioassay and absorption. The antisera employed were conventional polyspecific alloantisera, typing sera with restricted specificty and monoclonal hybridoma-derived anti-H-2k antibodies.The tumour has a low expression of the private specificty 31, which characterizes Kd molecules, and does not seem to express the private specificities of Dd, Kk and Dk molecules. It appears to express predominantly alien H-2-like antigens which are very similar to but not identical with normal H-2k molecules.

Notes: The immune response of SJL/J (H-25) mice to syngeneic reticulum cell sarcoma (RCS) tumour cells was investigated. The nature of tumour-associated antigens was examined by immunological and biochemical techniques. Reticulum cell sarcomas of different origins (spontaneous, transplantable and cultured cell lines) stimulate a strong syngeneic response measured by [3H]-thymidine incorporation. However, only the cell line were able to stimulate a syngeneic cell-mediated cytotoxic response. Further analysis of the syngeneic response revealed that RCS cells express inappropiate alloantigenic specificities on their surface. Thus, evidence is provided which demonstrates that RCS tumour cells carry antigens which cross-react with BALB/c (H-2d) and C57BL/6 (H-2b) alloantigens. The presence of inappropriate antigens in the in vitro lines was detected by cell-mediated cytotoxicity, cytotoxic antibody and immunofluorescence whereas antigens on in vivo lines and spontaneous RCS were detected primarily by immunofluorescence. Conversely, H-25 expression on the in vitro lines was detected only by immunofluorescence whereas the H-25 of the in vivo lines were detected by cytotoxic antibody. By these assays inappropriate antigens were not detected on normal SJL/J lymphocytes. Immunochemical analysis of the inappropriate antigens by two-dimensional gel electrophoresis was performed with SJL/J anti-BALB/c and monospecific anti-H-2Dd sera. These sera precipitated 45,000 MW molecules from BALB/c lymph node cells and from SJL/J in vivo and in vitro RCS tumour which appeared very similar. The sera also precipitated molecules from SJL/J lymph node cells which resemble the BALB/c and RCS molecules. Thus, RCS tumours express inappropriate alloantigens which may be cryptic on normal cells. The biological significance of these tumour-associated antigens in vivo is discussed briefly.

Notes: Serological and structural changes of surface markers involved in immune reactions may occur in human and murine tumour systems. Thus nine out of twenty-one human tumour cell lines and SV40-transformed fibroblasts differed from autologous lymphoblastoid cells or fibroblasts in their reactivity with HLA allonatisera. H-2 antigens isolated from the murine tumour cells 6C3HED and TP 1422, undergo structural changes. An alien HLA-B7 was detected in sera from two melanoma patients. The serologic activity on H-2 antigens was significantly increased in the serum and ascites fluid of tumour bearing mice. Additionally, human SV40-transformed fibroblatsts acquire receptors for monkey red blood cells and the murine lymphosarcoma cells 6C3HED express receptors for sheep red blood cells.

Notes: Rg and Ch typing was performed, by serum inhibition, on 145 families that had been typed for HLA/C4/BF/C2 with a view to assessing partial inhibition (p.i.) of anti-Rg/Ch and its haplotype associations. Rg p.i. was found predominatly with the C4A*3A*,2,B*QO homoduplicated C4 haplotype and BFF. The original type of Ch p.i. (Nordhagen et al., 1980) was closely associated with the allotype C4B 2, which also occasionally exhibited complete inhibition (c.i.), but this Ch p.i. was also found with the C4A*1,B*QO haplotype (Rittner et al., 1984a). The second type of Ch p.i. (Giles, 1984) was closely associated with the C4B 1 allotype most frequently in the haplotype C4A*6,B*1 but also with C4A*3,B*1. Both types of Ch p.i. are usually found with BF S. The present data indicate that the determinants of Rg and Ch are not directly related to any particular C4 allotype or extended haplotype.Further examples of C4A 1 with Ch and C4B 5 without Ch determinants have been detected and theoretical considerations are discussed as to how they might have arisen from unequal crossovers in homologous regions that result in hybrid protein molecules.

Notes: A comprehensive strategy for the systematic localization of all continuous antigenic sites within a protein has previously been introduced by this laboratory. The strategy consists of studying the immunochemical activity of a series of consecutive synthetic peptides that encompass the entire protein chain and the are uniform in size and in overlap at their N and C-terminals with neighbouring peptided. By application of this strategy to sperm whale myoglobin, we have been able to delineate the ontinuous sites of T cell recognition of myoglobin in three high responder mouse strains. Thirteen 17-residue peptides that encompass the entire myoglobin chain and overlap by five residues at both ends were synthesized, purified and characterized. The peptides were examined in vitro for their ability to stimulate lymph node cells from myoglobin-primed DBA/2 (H-2d), BALB/c (H-2d) and SJL (H-2s) mice as well as long-term cultures of myoglobin-specific T cells. Several regions of the moleculr (T sites) were founnd to stimulate myoglobin-primed lymph node cells and myoglobin-specific long-term T cell cultures. This strategy has enabled the localization of the full profile of dominant sites of T cell recognition in myoglobin for these mouse strains. Of these T sites, one region, residues 107-125, was clearly immunodominant in these strains and was found to coincide with the antigenic (i.e. antibody binding) site 4 of myoglobin. Also, other regions stimulated T cells and appeared to coincide with previously known antigenic sites. It is noteworthy that, in addition to sites recognized by both T and B cells, the protein has other sites which are recognized exclusively by T cells and to which no detectable antibody response is directed.

Notes: The IgM response to SRBC in a line of rats selected for high serum IgM was higher and lasted longer than that in a line of rats selected for low IgM, while the response to TNP-LPS was similar at its peak in the two lines but persisted longer in the ‘high’ line. The difference between the lines is therefore more likely to be at the macrophage and/or the T cell level than at the B cell level.

Notes: The relationship between immunoglobulin allotypes and leprosy was studied in 91 unrelated patients and 100 healthy controls from Vietnam. Twenty Vietnamese patients with tuberculosis were also typed for the Gm, A2m and Km allotypes. The results were compared with those from the healthy controls. No significant association was found for the allotypes G1m(z,a,x,f) G2m(n), G3m(g,b), A2m(1,2) and Km(1,3) between the two groups of patients and the controls. Heterogeneity in the distribution of G2m(n), G3m(b), A2m(2) and Km(3) was found when 60 polar-lepromatous (LL) patients and 27 borderlinetuberculoid (BT) patients were separated out of the 91 leprosy patients. In the LL patients there appeared to be a significantly higher frequency of G2m(n), G3m(b) and A2m(2) in coḿparison with the BT patients (P 〈 0.05). A significantly lower frequency of Km(3) was found in the LL patients in comparison with the healthy control group (P 〈 0.05). The frequencies of the Gm-A2m haplotypes and of the occurrence of the Km(1) and Km(3) in the Vietnamese population were calculated on the basis of the results in the 100 samples of healthy controls. The main haplotype is Gmaf;n;b (frequency 0.676), occurring with A2m1 (0.200), as well as with A2m2 (0.476)

Notes: All mice responding to the terpolymer GLA40 make GL, GA and GLA specific antibodies irrespective of their response to GL or GA alone. The mice displayed positive T cell proliferative responses against the homologous terpolymer, but no T cell responses were obtained with GL, which is non-immunogenic in mice. T cells from GLA immune mice, which are also responders to GA, such as mice of H-2 haplotypes a, b, d, k and r, could be stimulated by GA. T cells from GLA immune mice of H-2 haplotypes p and q which are non-responders to GA could not be stimulated by GA. On the other hand, T cells from H-2s mice immune to GLA and which are also responders to GA alone could not be stimulated by GA. Thus mice of H-2 haplotypes p, q and s recognize the terpolymer via ‘GLA' determinants alone, whereas mice of H-2 haplotypes a, b, d, k and r may recognize both GA and GLA determinants in GLA terpolymer.

Notes: Three monoclonal antibodies HU-11, HU-32, and HU-33, which recognize two distinct polymorphic determinants of human class II antigens, were found to cross-react with rat B cells carrying an RT1B region-associated specificity Ba-2.6. This is the first report demonstrating that xenoimmune antibodies raised against polymorphic determinants of human class II antigens are able to detect polymorphic determinants of class II antigens in a third party species.

Notes: In order to determine the specificity of antigen binding by double antigen-binding lymphocytes obtained from mouse bone marrow and spleen, three types of experiments were performed (1) a high excess of unlabelled antigen was tested for its ability to inhibit the binding of unrelated antigen to both single and double antigen-binding cells; (2) polyvalent anti-mouse immunoglobulin was assessed for its ability to inhibit antigen binding; and (3) the ability of one antigen to co-cap (or codistribute) with either another antigen or antiimmunoglobulin was studied to determine the spatial relationship of these components on the cell membrane. In order to study an adequate number of double ABC, these cells were enriched by using NIP-specific ABC as a starting population. The data indicate that double antigen binding occurs via independent immunoglobulin cell surface receptors which can be spatially separated from one another under appropriate capping conditions.

Notes: The relationship between the ability of female inbred mice to produce an anti-paternal humoral immune response to allogeneic multiparity and the genotype of the female and male strains has been investigated. Only three, all H-2b haplotype strains, were ‘responder’ strains and produced anti-paternal alloantibody which did not exhibit C'-dependent cytotoxicity. ‘Non-responder’ strains produced no alloantibody in spite of multiple pregnancies with H-2 and non-H-2 incompatible male strains. However, even responder strains did not produce alloantibody with all incompatible male strains. The absence of a response in a responder strain mated with a male strain differing at only the H-2 locus implicated a role for non-H-2 influences. A study of the specificityo of pregnancy-induced alloantibody suggested that this represented only a fraction of the total alloantibody population induced by conventional immunization in the same strain combination. It is suggested that in pregnancy only the anti non-H-2 humoral responses remain similar to those induced by immunization, whereas anti-H-2 humoral responses are either absent or are restricted to a fraction of the total foreign H-2 specificities presented. These observations are discussed in relation to the nature of the immunogenic stimulus in pregnancy and the expression of histocompatibility antigens on the feto-placental unit.

Notes: In order to determine the cell type responsible for the antigen-binding reaction in the bone marrow and spleen of mice, cells derived from pure in vitro derived colonies of neutrophils, eosinophils, macrophage-megakaryocytes and B lymphocytes were tested for their ability to bind fluorescent protein antigens. Only B lymphocytes bound antigen. An unexpectedly high percentage of bone marrow B lymphocytes (20%) bound a given antigen. This frequency was considerably higher than that found for spleen cells. As might be expected from such high binding frequencies, some cells bound two fluorchromated antigens when these are added together. As a direct test of the clonality of antigen binding to bone marrow B lymphocytes, whole colonies of B cells were tested for antigen binding of two non-cross-reacting protein antigens. The frequency of antigen-binding clones, including double antigen-binding clones, reflects exactly the frequencies observed for dispersed colony B cells and for in vivo derived Ig-bearing bone marrow B cells. The frequency of double antigen-binding colonies was equal to the product of the frequencies of the colonies binding each of the two antigens alone. No ‘mixed’ colonies containing single binding cells for each antigen were found. Thus, the ability to bind any two given antigens is a clonally distributed property of the bone marrow B lymphocyte population. Heterogenous receptors for multiple antigen binding on each cell are either randomly distributed among the B cell population, or homogenous antigen-binding receptors on each cell have a random chance of cross-reaction with the two antigens tested.

Notes: Morphological studies of metaphase chromosomes were done with rats derived from the BIL/1 strain, which has genes affecting growth and reproduction linked to the major histocompatibility complex by conventional Giemsa-trypsin staining and the results were compared to rat strains not carrying these defects. A subterminal-terminal centromeric polymorphism was detected in chromosome 3 [del 3 (pter → cent)] among the strains studied. Comparison of the G-banded karotypes of the rats carrying the defects with the karotypes of the otsher strains did not reveal any gross chromosomal abnormalities.

Notes: C4-coated Ch(a+) red blood cells (RBC) were used as indicator cells in a serum inhibition reaction of anti-Cha, for the determination of the Ch group of serum. This serological study, combined with electrophoretic studies of C4 in a family material, showed that the C4M haplotype product was associated with partial inhibition of anti-Cha.

Notes: Individuals with selective absence of IgG1 and IgG2 were discovered by testing for allotypes and isotypes of the respective sub-classes. These individuals were homozygous for sub-class deleted Gm-Am haplotypes, as shown by allotype studies in two families (Gm-;…;g;A2ml/Gm-;n;b;A2ml and Gm-;n;b;A2ml/Gm-;…;b;A2ml) and by a population study of New Guineans (Gm fa;-;b;A2m2). The individuals with IgGl sub-class deficiency showed elevation of IgG2, IgG4 and in particular of IgG3.Gene deletion can result from unequal crossing over which renders a complementary chromosome with a duplication of a sub-class gene. In one family, duplication of γ3 genes was observed to have happened in one of a twin pair. Quantitation of sub-classes in families with γ1- and with γ3-duplicated haplotypes did not show increased levels of the gene involved.

Notes: A hybrid cell line R3/41-10 was produced by fusing a mouse myeloma line with spleen cells from CBA/H mice which had previously been immunized with human peripheral blood lymphocytes (PBL) obtained from a single donor. Hybrid secreting antibodies to lymphocyte antigens were detected by assaying the culture supernatant for antibody binding to the donor PBL. Those lymphocytes binding more than ten times the background were cloned in soft agar and transferred to micro-culture plates (Limbro). Antibodies from some wells lost their binding activity. Antibodies from other clones, although retaining their binding capacity, were multispecific in cytotoxicity experiments, killing all the lympoid cells from a panel of normal donors; yet a third kind gave specific limited reactions. Supernatants from clones R3/41-10 and R3/41-13 gave concordant cytotoxic reactions killing 20-40% of the PBL. There was no cytotoxicity of Ig+ cells (B cells). Lymphocytes from seven out of fourteen normal donors reacted with the antibody. The specificity of the antibody produced by clone R3/41-10 was confirmed by absorption studies.The monoclonal antibodies (Mc+b) described in this communication are shown to react with a subpopulation of T lymphocytes of some individuals and not others, suggesting that they are detecting a polymorphic system of alloantigens like the Ly system in the mouse, provisionally designated HT-Ly. l.

Notes: Subcutaneous (s.c.) immunization of mice with viable allogeneic H-2 compatible spleen cells can induce a persistent state of delayed type hypersensitivity (DTH) to minor histocompatibility (H) antigens which can be evaluated with a footpad swelling assay. The importance of H-2 compatibility of the injected spleen cells with the recipient for (1) the elicitation of the DTH-reaction, (2) the induction of DTH-related effector and memory T cells and (3) the activation of T memory cells was examined with congenic mouse strains. Spleen cells sharing either the K or D region of the H-2 complex with the recipient could elicit strong DTH-reactions to minor H antigens, though somewhat less than did fully H-2 compatible allogeneic spleen cells. H-2 incompatible cells or cells only sharing I-region coded antigens elicited relatively weak, though significant, DTH-reactivity to the minor H antigens. Similar H-2 requirements for recipient mice were demonstrated in the immune lymphocyte transfer assay. Optimal induction of primary DTH and DTH-related T memory cells for minor H antigens also required H-2 identity of the immunizing cells and the recipient. To some extent, H-2 incompatible cells were able to induce primary DTH-reactivity and memory for minor H-antigens. The secondary DTH-reactivity was not or slightly dependent on the H-2 haplotype of the cells used for booster injection.It is concluded that the DTH-related effector cells are restricted in their recongnition of minor H antigens by K- or D-region-coded antigens. Presumably, macrophage processing of the allogeneic spleen cells after primary and secondary immunization accounts for the capacity of the minor H antigens to activate unprimed T cells and memory T cells when these antigens are presented on H-2 incompatible cells.

Notes: Difference in the amounts of H-2K.5 antigens present on erythrocytes was observed, using quantitative absorption method, among several B10 congenic strains. However, no such variation was seen on the lymphocytes and lung cells. The variability in the amount of this antigen on the erythrocyte surface was primarily dependent on the haplotype of the H-2K end in the B10 recombinant strains examined. This suggests that the regulator of H-2 expression on erythrocytes is in this region. Further genetic analysis confirmed that this regulation functions in the cis-position. Finally, the H-2K.5 antigenic activity was expressed on the reticulocytes of all strains tested, but appeared lower in the type 2 group indicating that the regulation begins early in erythropoietic differentiation and eventually results in a complete loss of detectable activity.

Notes: A two-step procedure employing gel filtration and anion exchange chromatography has been utilized to isolate LMW immunoglobulin from the horned shark, Heterodontus francisci. Light chains obtained by complete reduction and alkylation of the parent protein have been compared by several analytical techniques. Amino acid composition data implies a limited degree of variation in the light chains isolated from individual animals. Polyacrylamide gel electrophoresis of the CNBr digests of the light chains reveal indistinguishable banding profiles of the major peptides. Isoelectric focusing indicates limited heterogeneity in the light chain spectrotype and identity in the pI of the majority of bands detectable by staining. The suggested degree of structural similarity in the light chains of this phylogenetically primitive shark is discussed in terms of the evolutionary position of the species and current theories concerning the origins of structural diversity in immunoglobulins.

Notes: The mechanism of neonatally induced transplantation tolerance was studied in two mouse strain combinations involving differences at the D region of the H-2 complex only or at the same D region plus I-J subregion (including I-E, I-C, S and G regions). In the strain combination with the H-2D difference only, cells from tolerant mice proliferated markedly in the MLR assay when incubated with antigens tolerated in vivo, whereas the MLR reactions were negative in the combination with D plus I-J region disparities. In the latter combination cells from tolerant mice also did not respond to third-party antigens and their incubation with the tolerated antigens led to the suppression of cell proliferation. This non-specific suppression was absent in cells from tolerant mice in the strain combination, which differed in I-C, S, G and D alloantigens. Specific suppressor cells, which inhibited the development of cytotoxic cells, were demonstrated in tolerant mice of both strain combinations. The results show that, in addition to the specific suppressor cells induced by H-2K or H-2D alloantigens, non-specific suppressor cells induced by the I-J region disparity that may regulate the resultant activity against H-2D (and probably also H-2K) alloantigens are involved in transplantation tolerance.

Notes: We have used a combined serologic and structural approach to study the distribution of I-region associated (Ia) antigens in nine strains of inbred and partially inbred guinea-pigs. All of the inbred strains studied with the exception of strain 2 animals were found to share one or more I-subregions with inbred strain 13 animals. The BIOAD, R9, OM3, and BIOAC strains have the same I-region as strain 13 animals; the B/Lac strain has two subregions in common with strain 13, while the BIOB strain has a single subregion in common with strain 13. The availability of a number of different guinea-pig strains with well characterized major histocompatibility complexes should facilitate the continuing use of this species in studies of immunogenetics, transplantation, and tumour immunology.

Notes: Two-way and one-way mixed lymphocyte cultures (MLC) with Chinese Hamster peripheral blood lymphocytes were performed. Reduction of the culture surface and addition of 2-mercaptoethanol to the culture medium gives a marked enhancement of the reactivity of low numbers of lymphocytes in the MLC. Genetic analysis of the mixed lymphocyte reactivity (MLR) in the animals studied so far indicates that three or four different alleles are involved. Howerver, not all phenotypes could be traced. The samples of different stocks of Chinese Hamsters in Europe and the United States did not all share the same alleles and the samples were also different as regards the relative frequency of phenotypes. For DNA inhibition in stimulating cells mitomycin C treatment is less suitable than irradiation. Skin graft survival time was longer in animals which showed no MLR than in animals with a positive MLR.

Notes: The genetic basis of the restriction imposed on T cell mediating acquired antimicrobial resistance and delayed-type hypersensitivity (DTH) to Listeria in the rat was investigated. Sharing of MHC-coded genes between donors of sensitized T cells and antigen-stimulated recipients was both necessary and sufficient for efficient transfer of both resistance and DTH. Evidence to support this assumption was derived from experiments involving allogeneic transfers within major histocompatibility complex (MHC)-compatible strains and across MHC-barriers. Further support came from linkage studies with backcrossed rats and with the progeny of F1 rats mated with an unrelated strain. An unexpected difference in the compatibility requirements for effective transfer of DTH and resistance was noted in experiments involving the BI strain (formerly called B3). Thus, while B-region compatibility was obligatory for expression of DTH in recipients of sensitized T cells, considerable levels of protection could be transferred to either A-region or B-region compatible hosts.

Notes: The antibody response in mice to type I calf skin collagen is quantitatively determined by genes which map to the I region of the H-2 histocompatibility complex. The use of H-2 recombinant B10 congenic strains of mice reveals that a gene in the IA subregion and a gene to the right of the IA subregion affect responsivenss. To examine complementation patterns in the antibody response to collagen, five B10 congenic strains, each bearing an independent H-2 haplotype, were intercrossed to obtain nine hybrid strains heterozygous at the H-2 locus. In five combinations heterozygous progeny produced significantly greater antibody responses than those observed for the homozygous parental strains. Two low responder haplotypes, H-2k and H-2d, were shown to be qualitatively different. Mice of these haplotypes show a different dose-response pattern and a different phenotypic pattern of inheritance with respect to the high responder H-2b haplotype. Complementation effects found in F1 hybrid mice derived from H-2 recombinant parental strains indicate that high responsiveness, controlled by an IAb subregion gene, can be influenced by an interaction between an IAk subregion gene and an ICd subregion gene on different chromosomes. These data are consistent with the possibility that there exist two or more I region genes that have distinct functions and can interact to affect the levels of immune responsiveness.

Notes: A study of Gm allotypes in a Caucasoid family with hypogammaglobulinaemic probands, showed qualitative (unexpected or lacking Gm allotypes) and quantitative (increased or decreased Gm contents) abnormalities in many relatives. Part of these observations can be most probably accounted for by inheritance of a Gm1,17;5,28 haplotype, not described in Caucasians yet, and by an in vivo expression of latent Gm genes.

Notes: α-L-Fucosyltransferases were demonstrated in human saliva which catalyze the transfer of L-fucose from GDP-L-[14C]-fucose to oligosaccharides from human milk. An α-(1→4)-L-fucosyltransferase that synthesizes lacto-N-fucopentaose II and lacto-N-difucohexaose I from lacto-N-tetraose and lacto-N-fucopentaose I, respectively, was detected in saliva samples of Le(a-b+) secretors and Le(a + b-) non-secretors in which Lea substance was secreted. This enzyme activity was demonstrable neither in saliva samples of Le(a-b-) secretors nor non-secretors. An α-(1→2)-L-fucosyltransferase, that synthesizes lacto-N-fucopentaose I from lacto-N-tetraose, was detected in saliva samples from Le(a-b+) secretors which secreted H and Leb substances and from Le(a-b-) secretors which secreted only H substance. An α-(1→3)-L-fucosyltransferase was present in all saliva samples of different ABO and Lewis blood groups, irrespective of their ABH secretor status of the donors. The fucosyltransferases in saliva were activated by Mn++ or Mg++ ions, and were inhibited by ATP, GTP and EDTA. They had a broad pH optimun between pH 5.0 and 6.5.

Notes: Genes in the D region of the murine major histocompatibility complex, H-2, confer resistance to radiation-induced leukaemia virus. H-2D gene control appears to ensue at a step subsequent to virus infection, since elimination of virus infected cells does not become apparent until 3-5 weeks after virus infection. Nonetheless, almost immediately after virus infection, expression of H-2D-coded antigens is markedly elevated on the surface of thymocytes from resistant (H-2Dd) but not susceptible mice (H-2Ds or H-2Dq). This increased H-2D antigen expression triggers a vigorous cell-mediated immune response which probably plays a key role in resistance to leukaemia via elimination of virus-infected cells. A hypothesis is put forth to explain the induction of increased sythesis and expression of H-2D antigens. This hypothesis postulates that the oncogenic segment of RadLV bears a close resemblance to H-2.4, the private specificity for H-2Dd, allowing it to integrate at or near the H-2Dd murine gene. Subsequent to integration, the rates of transcription and translation are altered with a resulting increase in cell surface antigen expression. Other possibilities are also discussed.

Notes: When testing the serum of an individual anti-H-2 immunized mouse (B10 x A.SW)F1 anti-B10.M by the routine micro-lymphocytotoxicity test on lymph-node cells, unexpected antibodies were found. The most striking finding was that after absorption of anti-H-2.8 antibodies with B10.A(2R) (Kk) cells, antibodies remained which reacted with AKR, B10.AKM and B10.A V + mice while B10.A V-, B10.BR and C3H mice were negative. While all these strains share the Kk allele, only the positively reacting strains express high titres of infectious RNA turnover viruses. Unexpected reactions were observed also with H-2d, H-2j and H-2r cells and absorption experiments indicated two or three antibody populations.These reactions could be interpreted by two different possibilities: (1) anti-H-2 antibodies react with virus-altered H-2 structures; and (2) antiviral antibodies react with H-2 structures complexed with viruses. These possibilities should be taken into account when H-2 sera are tested on tumour or virus-infected cells.

Notes: Cytostatis of the H-2d tumour LSTRA by H-2-restricted effector lymphocytes was inhibited by antisera against H-2.4 and H-2.31 but not by antisera against public specificities or non-H-2 antigens. The unexpected reaction of the same effector cells against Gardner tumour (H-2k) was also shown to be inhibited by a combination of antisera against H-2.4 and H-2.31 but not by each antiserum used separately. The inhibitory capacity of these antisera was removed by absorption with B10.D2 but not with B10 lymphocytes. This indicated the presence of H-2d-like specificities on gardner tumour which could function as self-recognition structures in an H-2Kd and H-2Dd restricted system.

Notes: The specificity of he corss-killing exerted by cytotoxic T lymphocytes (CTL's) generated against H-2 region products was investigated in a 51Cr release assay on a panel of target cells from a number of different H-2 haplotypes.Their pattern of reaction shows that: (1) The target cells, expressing public specificities (H-2.28, H-2.1, H-2.5, H-2.3 or H-2.8) which should, theoretically, be recognized by the CTL's were killed, while those expressing no public specificities, recognized according to the H-2 chart by the CTL's wer not killed. (2) The CTL's generated against the H-2.28 specificity expressed on the D region products cross react with target cells expressing this specificity in their K region products and vice versa. The same phenomenon was observed with the H-2.1 specificity. These results provide evidence that public specificities are targets for CTL's. Antiserum reacting against the public specificity recognized by the CTL's was found to block the cross-killing, however, to the same extent as antisera directed against any specificity recognized by the CTL's was found to block the cross-killing, however, to the same extent as antisera directed against any specificity (private or public) expressed on the same molecule as the target determinant. Finally both the inhibition studies using anti-H-2 antisera and the direct cytotoxic assays showed that the public specificities of the H-2.28 family carried by the H-2.D and H-2.L molecules were recognized by different subpopulations of CTL's.

Notes: Three recessive lethal t haplotypes have been independently isolated from widely separated Chilean wild mouse populations. They do not complement one another, have similar transmission ratios and suppress recombination. We do not know at present if they belong to a previously described complementation group or if they define a new one.

Notes: Investigation of an informative family including two probands with male multiplex retinitis pigmentosa revealed that the putative disease susceptibility loci were not linked to those of HLA. In addition, analysis of immunological data obtained yielded evidence suggestive of X-linked inheritance of susceptibility to cell-mediated immune aberration in this family.

Notes: A recombinant haplotype between the class I and class II major histocompatibility loci of inbred miniature swine has made it possible to study the role of the corresponding antigens in in vitro and in vivo immune responses. Before examining the effects of selective SLA locus matching on allograft survival, it was deemed necessary to determine the corresponding effects on in vitro parameters of immunity. The results presented in this paper indicate that: (1) a difference at the class II loci was required for a maximal proliferative response which depended on the haplotype combination used; (3) in cell-mediated cytoxicity assays, the class I antigens were found to be the primary targets for lysis. Although optimal CTL generation required the presence of both class I and class II antigen differences, CTL were sometimes generated in the absence of a known class II difference. The weak, intermittent cytotoxicity seen in these cultures of class II difference. The weak, intermittent cytotoxicity seen in these cultures of class II matched cells could be augmented by the addition of third party, class I matched, class II mismatched, stimulator cells. Taken together, these data provide evidence for the conservation of function of antigen class between species. They also provide further evidence for the two-signal hypothesis of CTL generation. They, also provide further evidence for the two-signal hypothesis of CTL generation. Thus, MHC recombinant swine provide a model for the study of the effect of antigen class on in vitro and in vivo immunobiology.

Notes: Thymic immunosuppression in adult Xenopus laevis laevis, the South African clawed toad, is antigen-dependent and antigen-specific, but it is not genetically restricted. In this report, we show that combination in vitro with a thymus from an immunized Xenopus laevis laevis can suppress antibody production to a hapten from spleen fragments, if the spleen fragments are derived from a subspecies of Xenopus laevis or a species of Xenopus which shares the same diploid chromosome number (2N = 36). Some aspects of the evolution of thymic immunosuppression are considered in the light of these results.

Notes: Book Revies in this ArticleM. KIMURA: Molecular Evolution, Protein Polymorphism and the Neutral Theory.B. ROIZMAN: The Herpesvirus

Notes: New findings on the detection of alien histocompatibility antigens (AHA) on tumour cells are reviewed. Three points are discussed, namely: (1) isolation and molecular characterization of AHA; (2) frequency of expression of AHA by tumour cells, and (3) mechanisms of expression of AHA. Mechanisms of expressions of AHA are particularly discussed in the light of the recent advances in the understanding of the molecular genetic organization of H-2 antigens. Gene conversion involving coding or non-coding sequences might be responsible for the appearance of unexpected H-2 determinants on cancer cells.

Notes: Neonatal spleen populations have been studied for antigen-binding capacity in an attempt to determine if the frequency of double antigen-binding cells changes as the population matures. Just after birth, the frequencies of total ABC, double ABC and Ig-bearing cells were similar to the frequencies seen in the adult bone marrow. During the first week after birth, the proportion of total ABC and Ig-bearing cells rose sharply so that by 8 days after birth, these cells were about half of adult levels. The proportion of total double ABC, after a brief rise at day 1, remained constant throughout the test period. Thus, the proportion of total ABC which are doubles gradually decreases with age. Sedimentation velocity studies indicate that double ABC's tend to co-sediment with cells which require a period of maturation before they can respond to a thymus-independent antigen in irradiated hosts. Single ABC, on the other hand, tend to co-sediment with cells which are immediately responsive to antigen, or require a shorter maturational period before they become responsive to antigen. All of these data, taken together with our other work, suggest the possibility that multiple ABC are clonally-derived lymphocyte populations which become more restricted in their antigen-binding capacity as they mature.

Notes: Book reviewed in this article: Fish Pathogens and Environment in European Polyculture. Ed. by J. OLÁH. Recent Advances in Aquaculture Ed. By James F Muir and Ronald J. Roberts. Modern Methods of Aquaculture in Japan. By T. and H. Iklenone. Vol. 11 in Development in Aquaculture and Fisheries Science The Collection and Preservation of Animal Parasites. Compiled by M. H. Pritchard and G. O. W. Kruse. Technical Bulletin No. 1 of the Harold W. Manter Laboratory.

Notes: Abstract Dead lobsters from five lobster parks have been examined over a period of 5 months for the occurrence of Aerococcus viridans. Isolated strains of the bacterium were characterized biochemically and by means of intracellular protein patterns using thin layer isoelectric focusing in polyacrylamide gel. In a lobster park where a high mortality occurred, A. viridans was isolated from 29 of 33 dead lobsters. The bacterium was also isolated from 7 of 10 dead lobsters in a lobster park with low mortality. Probably, infection with A. viridans occurs more frequently in Norwegian lobster parks than hitherto suspected. Gram-positive cocci sharing many characteristics with A. viridans were isolated from some dead lobsters. These bacteria did not form tetrads, they were catalase positive and had different protein patterns from A. viridans.

Notes: Abstract The fate of intravascularly-injected colloidal carbon was followed using routine light and electron microscopical techniques. The colloid was observed in the gills, heart, liver, kidney, supraneural organ, blood and areas near the site of injection. There was no apparent build up, removal or translocation of the colloid over the time periods examined (1 h-12 days) except in the blood, where the carbon was cleared from the plasma after about 3 h. Carbon-containing granulocytes were, however, present in the blood up to 12 days post-injection. Most of the carbon was localized in the cavernous bodies of the gills, the intertubular tissues of the kidney and the supraneural organ where it was found within phagocytic cells containing melanin pigments. These cells are compared to the melano-macrophages of higher fish and their possible role in antigen trapping and antibody formation is discussed.

Notes: Abstract The possible mechanism of inactivation of the toxicity of Aeromonas salmonicida extracellular products (ECP) by normal rainbow trout serum was investigated using juvenile rainbow trout. ECP was prepared from culture supernatant by an acetone precipitation method. The ECP was incubated with normal rainbow trout serum at 20°C for 2 h, and the interrelationship between ECP proteolytic activity and immune complex-initiating, haemolytic complement activity (CH50) of normal serum against antibody-sensitized goldfish red blood cells was evaluated. When normal serum was incubated with increasing concentrations of ECP, the CH50 activity of serum decreased. The CH50 activity was completely abolished in serum treated with undiluted ECP. ECP treated with serum was administered to trout intraperitoneally to determine mortality. All the fish receiving untreated ECP (0.05 ml = 0.5 mg protein) alone died within 24 h. When ECP was treated with serum at 1:1 to 4:1 (serum: ECP) in volume a similar high mortality was produced. These inocula possessed high protease activity and no or low CH50 activity. However, mortality decreased and finally no mortality was recorded as ECP was treated with large volumes of serum (9:1 to 19:1). These inocula had lower protease activity and considerably higher CH50 activity. Fish receiving ECP treated with heat-inactivated serum at 19:1 showed 100% mortality. A serum: ECP inoculum derived from fish which had been administered lipopolysaccharide from Salmonella enteritidis and which possessed a low CH50 activity also gave a high mortality when used at 19:1. These results suggest that rainbow trout complement is implicated in the inactivation of toxicity of A. salmonicida ECP.

Notes: Abstract A detailed review of proliferative kidney disease (PKD) in salmonids is presented. The introduction places the disease in a historical context. The various aspects of the disease are described from the existing literature, together with latest findings from current research. Particular emphasis is given to epidemiology, aetiology and pathology.

Notes: Abstract Following an outbreak of botulism in the fish stock of a British trout farm in 1980, a survey of the residual contamination of the farm was carried out. Commencing in November 1980, the farm was examined monthly for one year after the disease had been eliminated. A nearby unaffected farm was examined as a control. Examination of pond muds showed that the affected farm remained contaminated throughout the year, counts of Clostridium botulinum type E ranging from 1-800/g compared with counts at the control farm of 0.1-0.7/g. Both C. botulinum types B and E were found in the control farm, either singly or together in individual samples of pond mud. Fish taken from the affected farm in May and August of 1981 showed a 77.1 % (s.e. ±7.7) contamination by type E. Examination of the environs of the affected farm yielded C. botulinum in 36 of 60 samples of mud and soil using enrichment culture techniques. Clostridium botulinum type E predominated in muds, occurring in 26 of 28 samples taken from the waters supplying and by-passing the farm. In contrast, of the 32 soil samples tested from around the farm, eight yielded type B whereas only three contained type E.

Notes: Abstract. Strains of Yersinia ruckeri, the enteric redmouth (ERM) bacterium, show greater serological diversity than previously supposed. A distinct sub–group of the otherwise homogeneous serovar I (Hagerman) strains includes the salmonid blood spot (SBS) bacterium. Two other new serological varieties, serovars IV and V, have been designated.

Notes: Books reviewed in this article:Theory and Management of Tropical Fisheries. Ed. by D. Pauly & G. I. Murphy.

Notes: Abstract. Unfertilized, water-hardened eggs, obtained from a coho salmon, Oncor-hynchus kisutch (Walbaum), with coelomic fluid containing large enough numbers of the kidney disease bacterium, Renibacterium salmoninarum, to make the fluid cloudy (∼4 × 109 cells/ml), were examined by cultural and histological methods to determine (a) whether an earlier finding of the pathogen in coho eggs could be corroborated, and (b) whether iodine (500 mg/l, in the form of povidone-iodine) was as effective at ridding them of the intra-ovum pathogen as it was at freeing them of the extra-ovum pathogen. The results confirmed that the pathogen does indeed occur within the egg (11–6%-15-l % of the eggs were infected in this manner), and suggested a location in the yolk for the pathogen. The findings also indicated that while iodine was very effective in killing the pathogen on the surface of the eggs it was completely ineffective in reducing the prevalence of the intra-ovum infections. The results point out the need for a method for treating eggs internally and confirm the inadvisability of using eggs from salmonids with cloudy coelomic fluid.

Notes: Abstract. The transmission of Yersinia ruckeri has been investigated in steelhead trout using asymptomatic carriers of the causative bacterium of enteric redmouth disease. It was found that unstressed carrier fish did not transmit the bacterium to recipient fish to cause either an epizootic or produce new carrier fish. However, when the carriers were stressed with heat, the bacterium was transmitted from the carrier to recipient fish producing a lower intestinal carrier state but no deaths. Examination of experimentally infected fish to determine the number of carriers among the survivors indicated that the frequency varied as a function of time following infection. When immunized fish, were challenged with Y. ruckeri they became temporary carriers of the bacterium for up to 3 days; but were not able to transmit the infection to healthy recipient fish.

Notes: Book reviewed in this article: Diversity and Adaptation in Fish Behaviour. By M. H. A. Keenleyside. Exotic Species in Mariculture. By R. Mann. Developments in Marine Biology. Vol. 1. Toxic Dinoflagellate Blooms. Ed. by D. L. Taylor and H. H. Seliger Phytoplankton and the Fish Kills in Loch Striven. Ed by P. Tett.

Notes: Abstract. Atkinsiella hamanaensis sp. nov. a marine mastigomycete isolated from ova of the mangrove crab, Scylla serrata (Forsskål), is described and illustrated. The fungus grew over a temperature range of 15–32°C, with an optimum of 29–32°C. Its growth was observed in peptone-yeast extract glucose broth containing 1–5% NaCl, with optimum growth at 2–3% NaCl concentration. At 6% or more NaCl concentration, growth was inhibited. Its pH tolerance ranged from 4 to 9.

Notes: Abstract. Broth filtrate of a Vibrio species, isolated from spontaneously occurring epizootics among cultured oyster larvae, Crassostrea virginica (Gmelin), was shown to contain an exotoxin. Bioassays demonstrated that LC50 value was less than 46.6 μg of toxin/l culture of oyster embryos. This quantity was produced by 2.9 × 109 colony-forming units. Purified toxin showed neither proteolytic nor amylase activity; it did, however, demonstrate bacteriostatic capability. Studies showed that the toxin was heat-labile and that exposure to heat-produced toxoid had a beneficial effect on oyster embryonic development. Data revealed that although toxin was inactivated by heating, the bacteriostatic capability of the metabolite was not lost. Molecular weight of the toxin was estimated to be 68000.

Notes: Abstract. Flagellates infesting the gills of freshwater fish are reviewed and Cryptobia branchialis Nie is differentiated from them. Gill filaments of two species of fish heavily infested with C. branchialis were examined with the transmission electron microscope. The flagellates are attached to the surface of the gill epithelium by their recurrent flagella. A prominent ridge on the flagellar membrane contacts the host cell membrane, forming a zone of gap junction and leaving a space of about 10 nm between the two membranes. The flagella do not penetrate into the epithelial cells. No other modifications of the cell membrane could be detected and the host cell reveals no signs of damage. Assumptions on the pathogenicity of C. branchialis are discussed in view of ultrastructural findings, which suggest that this flagellate is non-pathogenic and ectocommensal.

Notes: Abstract. Gas bubble disease in larval striped bass, Morone saxatilis (Walbaum), was characterized by overinflation of the swimbladder and the formation of intestinal bubbles. This accumulation of gas hindered normal swimming of the fish and in extreme cases resulted in floating fish. As gas accumulated in the gut, the inner mucosal lining was compressed down into a cuboidal epithelium and in more advanced cases nearly the entire digestive tract was reduced to a squamous inner epithelium surrounded by a thin serosa. A significant increase in swimbladder volume was observed at total gas pressures as low as 102.9% and mortality was increased at 105.6-106.0%. The older larvae (30-day-old) were less sensitive to gas supersaturation than 10- to 19-day-old larvae.

Notes: Abstract. A rainbow trout with a malignant lymphoma of probable thymic origin showed direct spread of the tumour to the gills, metastases to the liver and head kidney, and the presence of abnormal circulating lymphocytes. The cells of the tumour possessed plasma membrane immunoglobulin detectable by immuno–fluorescence. Examination of the tumour cells by transmission electron microscopy revealed no evidence of virus associated with the cells. Infectious viruses could not be detected in the tumour tissue, and the tumour was not transmissible by injection of live tumour cells into young rainbow trout. Attempts to establish the tumour in long–term culture were unsuccessful.

Notes: Abstract. The sensitivity of the immunoperoxidase (IP) and fluorescent antibody (FA) techniques applied to frozen sections of organs of carp infected with spring viraemia virus (SVCV) was similar, both in respect of the intensity of the reaction and in the detection rate of the antigen. Seven days after a waterborne infection both methods detected the antigen in the kidneys and to a lesser extent in the liver and spleen. Quantification of virus gave a titre of 102.8 to 105.5 TCID50/g of kidney, whereas the agent could not be isolated from liver and spleen tissue. In contrast, at 241/2 days post–infection the viral antigen could be readily detected in liver and spleen but only occasionally in the kidneys using the IP and FA techniques. At this time, liver and spleen showed infectivity titres of 103.8 to 107.2 TCID50/g tissue, whereas the kidneys were found to be free of infective virus. It is concluded that using the IP and FA techniques SVCV can be detected most frequently in the kidneys from 7 to 14 days post–infection and in the liver and spleen from 14 to 24 days post-infection.

Notes: Abstract. A bacterin for immunization against bacterial kidney disease of salmonid fishes caused by Renibacterium salmoninarum is described. Cultures were grown in Evelyn's KDM2 medium containing 10% calf serum in a fermenter under the following conditions: pH 7.2, 15°C, 800ml/min air, 200 rev/min agitation and 5–15 days of incubation. Possible substitutes for calf serum were 10% horse serum 0.15% starch and leptospira medium. The bacterins were inactivated with 0.3% formalin and no adjuvants were used. Other tests evaluated pH-lysed bacterin, 50% concentrated bacterin and 50% concentrated pH-lysed bacterin. Juvenile rainbow trout, salmo gairdneri Richardson, were vaccinated either by intraperiotoneal (i.p.) injection, 2 min immersion or 2-step hyperosmotic infiltration. Fish were held from four to six weeks at 11°C, then challenged by i.p. injection with the homologous virulent bacterium. Fish died from days 19 to 40 after challenge. The best preparation was pH-lysed bacterin given by a single i.p. injection; hyperosmotic and immersion vaccination were not effective. Typically when 80% or more of unvaccinated controls were infected as detected by Gram stain, 10% or less of the vaccinated fish were infected.

Notes: Abstract. This paper reports for the first time the application of the direct negative staining technique for the rapid confirmation of the viral nature of blenny‘piscine erythrocytic necrosis (PEN). The virus in negatively stained preparations has an internal core structure which appears in exact opposite contrast to that seen in thin sectioned virus. The resultant images from both negative staining and thin sectioning suggest that the organization of the core is unique amongst intracytoplasmic deoxyriboviruses (ICDVs).

Notes: Abstract. Electron microscopical and histopathological investigations were carried out on the anterior kidney often 1-year-old farmed rainbow trout naturally infected with the external signs of viral haemorrhagic septicaemia (VHS). The histopathology of the liver and the haematology of peripheral blood were also examined. The presumptive virus of VHS was directly demonstrated for the first time in the anterior kidney.All diseased fishes showed necrotic changes in the kidney and the liver. The melano-macrophage centres of the anterior kidney showed a partial destruction accompanied with a loss of melanin. A progressive destruction of the erythrocytes was observed. The liver exhibited moderate to extensive necrosis (karyorrhexis and karyolysis with lymphocytic invasion in the necrotic areas). Neither intranuclear nor intracytoplasmatic inclusions were observed. In the peripheral blood diverse stages of erythrocytic degeneration, amitoses of the erythrocytes and a five-fold increase in lymphocyte numbers were observed.