ALBERT

Notes: Five human teratoma cell lines have been characterized for the presence of a certain number of marker antigens whose presence or absence has been shown to be characteristic of mouse embryonal carcinoma (EC) cells. Four out of the five lines have been shown to respond to at least some of the criteria associated with murine EC cells even though only limited in vitro differentiation could be demonstrated. The significance of certain unusual marker antigen combinations present on the cell line Tera I and its clones and so far unobserved for the murine model is discussed. The observation in Tera I populations of cells carrying simultaneously both the F9 and β2-microglobulin or HLA antigens, suggest that the human cell lines may represent a novel material for the study of mammalian differentiation.

Notes: Principles of Gene Manipulation. Studies in Microbiology

Notes: C4 is composed of two tightly linked genes (C4A and C4B) lying within the major histocompatibility complex of chromosome 6 that can be demonstrated by agarose gel electrophoresis. Seven alleles and five alleles at the C4A and C4B loci, respectively, were detected in 169 black individuals from the southeastern United States. Furthermore, the phenotypic frequencies of C4A6, C4A5, C4A4, C4B4, C4B3, and C4BQO were significantly different between black and white Americans.

Notes: Two B complex genotypes, B1B1 and B19B19, of outbred line S1, were tested for low and high immune response to GAT, from which four recombinants were recovered: B1B1 GAT-hi and lo, and B19B19 GAT-hi and -lo. Also included in the study were birds of B2B2 genotype with an intermediate level of immune response to GAT.A total of 225 birds of these groups were challenged with the Bryan strain of Rous Sarcoma virus subgroup C, RSV (RAV-7), by inoculation into the wing web at five weeks of age. The B1B1 genotype had the lowest percentage of regressors (17.6%), B19B19 had the highest (42.2%), and the B2B2 genotype was intermediate (23.7%). Combining the results of GAT response over the B1B1 and B19B19 genotypes, 14.0% of GAT-lo and 37.8% of GAT-hi regressed their tumours, respectively. The highly significant (P ≤ 0.01) difference between the combined GAT-hi and -lo groups would suggest that the Rs locus controlling tumour regression induced by the subgroup C virus is closely linked to the region controlling immune response to GAT, but the data also provides evidence that the B-F region of the B complex also plays an important role in RSV-induced tumour regression.

Notes: Surface immunoglobulin on spleen cells from NZB and NZB/W mice and congenic mice bearing the nude or X-linked immune defective (Xid) gene was examined by flow microfluorometry with regard to both the frequency of positive cells and density expressed on the cell. These data indicate that although the frequency of unseparated sIg+ B lymphocytes is equivalent among all of these groups of mice, the densities of sIgM and sIgD are different. Spleen cells from these mice were also separated by free-flow electrophoresis and analyzed in a similar manner. This analysis demonstrated the absence of a subpopulation of B lymphocytes with a low electrophoretic mobility and low expression of sIgM. These studies suggest that maturational and/or activation states of the B cells in mice bearing the Xid or nude genes are different from those seen in the parent strains of mice. Such alterations in cell-surface antigens correlate with differences in the natural history of immunopathology of the autoimmune disease in these congenic colonies of New Zealand mice.

Notes: Fifty-five Caucasoid patients with polymalgia rheumatica (PMR) or giant cell arteritis (GCA) were immunoglobulin (Gm) allotyped for this study. Forty-four of these patients had been previously HLA-A,B,C and DR locus allotyped. The incidence of the immunoglobulin allotypic marker Glm(2) was significantly increased in the GCA group (50.00% v. controls 18.75%, P= 〈0.01). There was a similar but insignificant rise of this Gm marker in the PMR group (27.24% v. 18.75%, NS). The increase in Glm(2) in the GCA group was not accompanied by a corresponding rise in the number of people homozygous for Glm(2), i.e., all the increase could be attributed to patients with the Glm(1,2,3): G3m(5,10,21) phenotype.

Notes: The strains B 10.S(7R), B 10.S(23 R) and B 10.S(24R), all thought to be genetically identical, differ in levels of susceptibility to infection with Trichinella spiralis. In a series of nine independent experiments, B 10.S(7R) was shown to be more susceptible than the other two strains. In another series of seven experiments, the strain B 10.A(18R) was shown to be more susceptible to infection with T. spiralis than the strains B 10.S(21R) or B 10.BAR-5, all of which were thought to share common H-2 alleles. These results indicate that a gene mapping between the S and D loci influences susceptibility to infection with T. spiralis. Typing of these strains for Qa and Tl loci rule out the possibility of a double crossover accounting for the differences observed. The new gene is designated Ts-2. Previously published data have also been reinterpreted and another gene Ts-1 is shown to be associated with the Aβ locus. When the d allele is expressed at the Ts-2 locus, strains of mice expressing s, q, f or b alleles at Ts-1 are rendered more susceptible to infection.

Notes: IgG and IgA heavy chain allotypes were determined in the sera of 483 Caucasian Type 1 diabetes patients and 503 Caucasian healthy controls. There was no significant difference between patients and controls neither on the level of Gm phenotype frequencies nor on the level of Gm three-locus and two-locus haplotype frequencies. A selective IgA deficiency was found in 14 patients (2.9%) but in none of the control individuals (P〈10-4).

Notes: Gm allotypes were detected and quantitated by radioimmunoassay (RIA) in paired serum and CSF samples from patients suffering from various neurological diseases. Of 115 patients with neurological disorders (65 MS and 50 others), seven subjects displayed one or two allotypes in their CSF which were absent in serum. The Gm phenotype in the patient's serum allowed us to infer the genotype without the need of familial data. A comparison of the regression curves obtained in RIA from the unexpected allotype in CSF and the counterpart in a normal serum pool argued for an identity of the Gm antigen carried by both inhibitory molecules. The unexpected allotype(s) in CSF can be considered as the product of a latent Gm gene which may be activated by either immune perturbations due to the disease per se or some particular immune regulations in the central nervous system.

Notes: Book reviewed in this article:N. Catsimpoolas; Cell Analysis. Plenum Publishing CorpJ.W.Shay: Techniques In Somatic Cell Genetics

Notes: Immunoprecipitaon studies of the rhesus monkey major histocompatibility system have shown that the RhLA-DR locus codes for class II antigens with molecular features that are homologous to the class II antigens coded for by the human HLA-dR locus, The products of another alloantigenic RhLA-linked locus of the rhesus monkey, called ‘48’, is provisionally characterized as a class I system.

Notes: The relationships between the antigens recognized by four monoclonal anti-human ‘Ia’-like antibodies were investigated using sequential immunoprecipitation and capping techniques. Two of the antibodies were ‘monomorphic’ and have previously been shown to recognize epitopes in which carbohydrate residues are involved, whereas the two ‘polymorphic’ antibodies recognized protein-defined epitopes—one of these epitopes being present on MB+DR- molecules. In the absence of an indisputable anti-DR monoclonal antibody, it was not possible to conclusively verify which ‘Ia’-encoded antigens were detected by the anti-‘Ia’-like monoclonal antibodies. Nevertheless, several firm conclusions could be drawn: (a) so-called ‘monomorphic’ antibodies do not necessarily react with all ‘Ia’ molecules encoded by a single locus—from the results using the two monomorphic antibodies, B5.1 and 3F1.1, described herein, two populations of antigens being B5.1+3F1.1+ and B5.1+3F1.1- were identified; (b) cross-reactivity of a polymorphic determinant expressed on antigenically-separable ‘Ia’ molecules was noted—using the two polymorphic antibodies, 26.1 and F5C9, molecules which were 26.1+F5C9+ and 26.1-F5C9+ were identified; and (c) the data clearly point to the existence of at least two loci coding for ‘Ia’-like antigens (one of which may or may not be the HLA-DR locus). Given that polymorphisms can now include protein- and carbohydrate-defined epitopes, that cross-reactions occur and that the definition of DR itself by monoclonal antibodies is not clear, the complexity of the human ‘Ia’ antgens is apparent.

Notes: We report HLA genotypes in four familial cases of Hodgkin's disease (HD), Nodular Sclerosis (NS) histological subtype, where all patients showed B18 antigen. This finding, although statistically not supported, confirms the possible correlation between HD and B18 antigen which carries a high relative risk in international data.

Notes: Previous studies from this laboratory have resulted in the determination of the antigenic structure of sperm-whale myoglobin (Mb). In the present work, we have investigated the fine specificity requirements for T-cell recognition of one of the Mb antigenic sites (antigenic site 5). The antigenic site (peptide 145-153) and seven progressively longer peptides, increasing in length stepwise by two residues at a time, up to 22 residues in length (peptide 132-153), were synthesized. In addition, four truncated peptides were synthesized with intentional deletions at Tyr- 151 and Ala- 144. The T-cell recognition of these purified synthetic peptides was examined here in detail in three strains of mice (BALB/cByJ, B10.D2/n and SJL/J). Mb-primed mice afforded T-cells which proliferated to smaller peptides (two or four residues longer than the site; i.e. peptides 145-153 and 143-153) and more so to the longer peptides 135-153 and 132-153 and to Mb. No response was obtained to the truncated peptides, thus underscoring the fine specificity T-cells. No response was obtained also to intermediate-sized peptides. The latter result, due to an unfavourable mode of folding, suggested a conformational dependency in T-lymphocyte recognition.

Notes: Serum blood samples from 563 of the total 700 Nganasans, members of the isolate in the northern-most part of Siberia were tested for G1m, (z,a,x,f), G2m (n), G3m (g,b0,b1,b3,b5,s,t), and km (1) allotypic determinants. Additionally, 78 Yenisey Samoyeds (Entsi) who are the Nganasan's western neighbours were studied. Both populations are remarkable for high frequency of ‘Northern Oriental’Gm (za;.;b0b3b5st) which appears to be the most frequent haplotype in the Nganasans (0.486), and is the second frequent in Yenisey Samoyeds (0.276). The Gm (f;b) generalized haplotype which used to be considered as an indicator of Caucasian gene flow occurred in the Nganasans in the very low frequency of 0.008, versus 0.045 revealed in adjoining Yenisey Samoyeds. Both populations also differ in the frequency of Km1 which is two times lower in the Nganasans (0.048), than in Yenisey Samoyeds (0.103). When segregation ratios for the Gm locus were inspected in 67 Nganasan families, no apparent deviations from Mendelian expectations, and no recombinant phenotypes were observed.

Notes: Genetic control of PQ prolongation of the electrocardiogram (ECG) in the mouse, immunized with killed group A streptococci, was studied by using various congenic mice. Mice of H-2a, H-2k and H-2f haplotypes showed high frequencies of PQ prolongation, while haplotypes of H-2b, H-2d and H-2s showed low frequencies of PQ prolongation. Studies using various recombinant mice revealed that at least one immune-associated (Ir) gene mapped in the left side of the I-B subregion. High responsiveness of F1 hybrids of H-2b and H-2d, as well as B1O.A(5R) and B10.A(3R), suggests the existence of a complementing gene. In addition, the differences between C3H and CKB, as well as differences between C3H.SW and CWB, indicate that another Ir gene maps in the immunoglobulin heavy chain (Igh) coding loci. Repeated injections of anti-I-J or anti-I-A antisera also modified this PQ prolongation. These results suggested that both the major histocompatibility complex (MHC) and immunoglobulin (Igh) loci seem to be playing important roles in the pathogenesis of PQ prolongation.

Notes: The growth and reproduction complex contains recessive genes (grc) which influence body weight and gonadal development. Homozygous males are sterile, and they have an arrest of spermatogenesis at the primary spermatocyte stage. Homozygous females are fertile but have a reduced reproductive capacity. The data presented in this paper show that the latter defect is associated with a decrease in the relative number of secondary ovarian follicles and an increase in the number of atretic follicles. This finding indicates that most of the primary follicles do not mature properly. Thus, the genetic defect in gametogenesis controlled by the grc appears to occur at the same stage of development in both females and males.

Notes: Serum IgG (7S) levels differed significantly for chickens from 10 different inbred lines. Within lines differences between B blood groups were statistically significant.The genetic control of serum IgG was further examined using birds from B complex haplotypes marked at the B locus and the Ir-GAT locus. Birds from each of five subgroup haplotypes (B1B1Ir-GAT-Lo and -Hi, B19B19Ir-GAT-Lo and -Hi, and B2B2Ir-GAT intermediate) were tested for levels of serum IgG at 3, 6, 9, and 21 weeks of age. The rate and level of IgG reached in the serum was more than two-fold greater in the GAT-Hi birds than in the GAT-Lo. The Ir region of the B complex exerts some control over the ontogenesis of IgG, though most of the genetic variation seems not to be B complex associated.

Notes: Dorso-ventral vaginal septa were observed in congenic mouse strains. Strain C57BL/10Sn, H-2b, had the highest frequency, 22%; B10.A/SgSn, H-2a, had the lowest frequency, 4%, P 〈0.001. The frequency was about 17% in the F1, females. Septa were found in 16% and 15% of the B10.D2/nSn and B10.BR/SgSn strains, respectively, indicating that at least two loci within the H-2 complex influence its frequency.

Notes: The phylogenetic distribution of antigens present on human lymphocytes was investigated by incubating human or simian cells with murine anti-human monoclonal antibodies and then determining the level of reactivity with a radiolabelled anti-murine IgG reagent. The monoclonal antibodies used were specific for a T-cell antigen, lymphoid and lymphoid:myeloid antigens, Ia antigens, and β2 microglobulin. The cells examined included B- and T-lymphoblastoid cell lines and fresh peripheral blood lymphocytes separated by sheep erythrocyte rosetting into T-cell and non T-cell fractions. Results of these studies showed that the antibodies gave complete cross-reactivity with gorilla and chimpanzee cells while B-cell lines of orangutan origin had lost lymphoid and β2 microglobulin markers. Gibbon cells and cells of Old World and New World monkeys reacted strongly only with monoclonal antibodies against Ia antigenic determinants. These Ia antigens were found on the non T-cell fraction of fresh peripheral lymphocytes, on B-cell lines and on some virus induced T-cell tumour lines. Immunoprecipitation analysis using the anti-Ia antibodies showed a degree of molecular diversity on owl monkey and marmoset cells compared to the Ia antigens associated with human cells.

Notes: C3H/HeN x NZB/BIN F1 mice were primed and challenged with DNP-Ficoll, a thymic independent (TI-2) antigen. They developed strong IgM and IgG carrier-specific memory responses: DNP-pneumococcal polysaccharide or DNP-haemocyanin challenge did not elicit memory responses. The IgG response component appeared to be inherited dominantly from the C3H/HeN parent. The IgM component resulted from genetic enhancement.

Notes: The effect of incompatibility for Mls determinants was studied in lethal graft-versus-host reaction (GVHR) in the mouse. GVHR was induced in adult recipients of the following H-2k strains: (AKR x B10.BR)F1 (MlsB/Mlsb); (C3H x B10.BR)F1 (Mlsc/Mlsb); (CBA/J x B10.BR)F1 (Mlsd/Mlsb) and (CBA/H x B10.BR)F1 (Mlsb). Recipient mice were heavily irradiated and grafted with bone marrow and spleen cells from H-2 compatible B10.BR (H-2k, Mlsb) or H-2 incompatible B10.D2 and B10 donors were normal, while those from B10.BR donors were either normal or pre-immunized against the recipient strains. In all experiments the survival of recipients with Mlsa/Mlsb and Mlsd/Mlsb phenotypes, and only in one experiment of those with Mlsc/Mlsb phenotype was greater and/or the survival time longer than that of recipients expressing only Mlsb. However, late deaths (〉 120 days post grafting) observed after grafting of normal B10.BR cells were more frequent in Mlsd/Mlsb than in Mlsb strains. On the other hand, when B10.BR donor cells were pre-immunized against H-2k compatible (AKR x B10.BR)F1 (Mlsa/Mlsb) or (CBA/J x B10.BR)F1 (Mlsd/Mlsb) strains, the survival time of H-2 incompatible (B10 x B10.BR)F1 (H-2b/k, Mlsb) recipients was longer than when donor cells were pre-immunized against (CBA/H x B10.BR)F1 (Mlsb) strain. We conclude that donor incompatibility for Mlsa or Mlsd or donor-pre-immunization against Mlsa or Mlsd exerts a protective effect on lethal GVHR developed across non-H-2 or H-2 barriers; the protective effect of Mlsc is less efficient or absent. The Mls-induced protective effect shows the following properties: (a) efficiency in vivo correlates with the capacity of the corresponding alleles to stimulate an in vitro MLR; (b) is efficient in either primary or secondary response to other minor antigens; (c) is not H-2 restricted; (d) is nonspecific; (e) disappears late after grafting; (f) with respect to the genetic background, the early protective effect is followed, late after grafting, by an opposite effect which increases the mortality, suggesting that M/s locus determinants are capable of activating several cell populations with different biological functions.

Notes: A long-term in vitro grown T cell line derived from a cotton-topped marmoset (Saguinus oedipus)) infected with Herpesvirus saimiri was found to share surface antigens with human amplifier T cells and to augment the capacity of human B cells to secrete immunoglobulin. This is the first demonstration of T/B collaboration across such a large phylogenetic barrier and might have interesting implications for understanding the nature of molecular interactions mediating cell/cell cooperation.

Notes: The segregation of genes controlling ECIA susceptibility and the level of immune response to native calf type II collagen were determined in the F2 progeny of matings between WF (RTIu/u; ECIA-susceptible; high responders) and LEW.B3 (RT1n/n; ECIA-resistant; low to intermediate responders). RT1n/n F2 progeny showed resistance to ECIA, low skin test reactivity to type II collagen and intermediate levels of IgG anti-collagen antibodies (-log2 of 6.2 ± 2.6; mean ± SD, n= 10). RT1u/u and RT1u/u F2 progeny were susceptible to ECIA and were high responders to type II collagen by skin testing and IgG antibody titres (-log2 of 12.1 ± 1.3, mean ± SD, n= 26). Although all rats that developed arthritis were also high responders to type II collagen one group of immature F2 progeny, RT1u/u and RT1u/n, showed high anti-collagen immune responses in the absence of detectable arthritis. The data indicate that genes linked to RT1.A control susceptibility to ECIA and at least part of the immune response to native calf type II collagen in WF and LEW.B3 rats.

Notes: Mice bearing the I-A subregion mutation bm12 were immunized and challenged with the α-subunit of human adult haemoglobin. Under conditions in which parental B6/Kh mice respond, B6.C-H-2bm12 mice are inhibited nearly 100% in their ability to respond to challenge to the α-chain of haemoglobin. D2.GD mice which express a variant Ae (Eβ) polypeptide of the I-E subregion can respond as well as B10.D2 mice to both subunits (α- and β-) of haemoglobin. These observations as well as other genetic mapping data confirm the I-A mapping of α-chain-specific Ir genes and extend the genetic fine mapping to the Aβ gene within the I-A subregion or a combinatorial Ia determinant generated by an interaction of Aα and Aβ. In addition they implicate the Ia.8 specificity in determining immune responsiveness to α-chain determinants.

Notes: Electrophoretic and serologic investigations of C4 in serum samples from a family material, showed that a weak, or partial, inhibition of anti-Rg sera, was an inherited property of some of the C4 haplotype products. The weak Rg activity was strongly, but not absolutely associated with the C4 FI and C4 I haplotypes.

Notes: Mouse oocytes at the dictyate stage were tested for the presence of H-2 antigens and Ia antigens, by means of indirect immunofluorescence. Oocytes reacted positively with NIH alloantisera, but were negative with anti-Ia region alloantisera. Results with monoclonal antibodies indicate very low antigen density on oocytes.

Notes: A Friend virus-induced tumour of BALB/c (H-2d) origin, HFL/d, was examined for the expression of alien H-2 antigens. The alloantigens on HFL/d were typed by generating CTL in primary MLC with HFL/d as stimulator and measuring reactivity to targets with known H-2 antigens, and confirmed by assessing recognition of HFL/d targets by CTL generated in primary MLC with stimulators expressing known H-2 antigens. Potential cross-reactivities between alloantigens were analysed by cold-target inhibition experiments. BALB/c cells stimulated with HFL/d lysed H-2b targets, and BALB/c anti-H-2b CTL lysed HFL/d; analysis with recombinant haplotypes demonstrated both H-2Kb and H-2Db alien antigens on HFL/d. C57BL/6 (H-2b) cells stimulated with HFL/d recognized H-2Kd, H-2Dd, and an additional determinant unique to HFL/d. (BALB/c x B6)F1 cells also recognised a unique HFL/d determinant not of H-2b or H-2d origin. These unique determinants, which induced a strong cytotoxic response in primary MLC, were not shared by BALB/c or B6 tumours induced by cross-reactive FMR viruses. Thus, HFL/d expressed the K and D antigens of its strain of origin, two typed alien H-2 antigens, and at least one other untyped antigen which may represent an additional H-2 determinant. These studies further demonstrate the utility of examining the reactivity of CTL generated in primary MLC to probe for the presence of alien H-2 antigens.

Notes: Properdin factor B is a genetically-controlled polymorphism that can be demonstrated by agarose gel electrophoresis. The Bf gene frequencies were determined for 194 blacks from the south-eastern United States. The frequencies for Bf-F, Bf-S, Bf-F1, and Bf-Sl are 0.626, 0.327, 0.034, and 0.013, respectively. The south-eastern blacks have similar Bf frequencies to the South African blacks; however, both these groups are different from blacks from Boston. The frequency differences between south-eastern blacks and Boston blacks may be due to racial admixture differences found in the two populations.

Notes: Fifty-seven members from ten families in which one parent and at least one child have asthma were studies with dilutional skin tests and RAST to grass pollens after determination of HLA haplotypes. We found no direct evidence for linkage of a hypothetical asthma locus with HLA or for a significant association of asthma with HLA haplotypes. Linkage between the HLA loci and a gene or genes which allow for the expression of clinical asthma could neither be proven nor disproven due to the small sample size. All of the asthmatic children had positive dilutional skin tests and RAST, suggesting that atopic asthma may be genetically controlled by the HLA chromosome (chromosome 6). Nonetheless, determination of the histocompatibility antigens can increase the value of predictive risk analysis for asthma. Such a determination may be important in the early identification of a child born to a family with atopic asthma.

Notes: T. OHTA; Evolution and Variation of Multigene Families.

Notes: The distribution of Glm(f, z, a and x), G3m(b0, b1, b3, b5, c3, c5, g, s and t), A2m (1 and 2) and Km(1) allotypic determinants in a series of Spanish blood donors. The following results were obtained: Gmz,a;gAm1= 0.199; Gmz,a;gAm2= 0.019; Gmz,a;bAm1= 0.081; Gmz,a,x;gAm2= 0.005; Gmf;bAm1= 0.677; Gmz,a;bAm1= 0.019 and Km1= 0.096. The results are similar to those reported for adjacent Basque and Portuguese populations. The origin of the Gmz,a;bAm1 haplotype is unknown.

Notes: Serolgoical and immunochemical assays detected antibodies to human la-like antigens in five out of five rabbit antisera to cultured human B lymphoid cells but did not detect them in five bleedings from two rabbits immunized with human peripheral blood lymphocytes. These results, which may reflect differential immunogenicity of la-like antigens from different cells, indicate that the presence of anti la-like antibodies in heterologous antihuman lymphocyte sera is not a general phenomenon as reported recently for animal model systems.

Notes: The murine Ly-4 locus is further described with the definition of both Ly-4.1 and 4.2 specificities and the demonstration of linkage of H-3 on Chromosome 2. As has been found for several other Ly-specificities, both Ly-4 specificities are absent from the thymus, but are found on peripheral T and B cells, with B cells having the greatest antigen expression. The expression of both Ly-4 specificities was influenced by an H-2-linked gene, mapping in or near the H-2D locus.

Notes: The factorial correspondence analysis, which is a descriptive statistical method, was used to study the preferential associations between locus HLA-A, B, C and Bf alleles. This method does not limit the number ofloci that can be studied and must therefore be considered as an interesting method of studying haplotypic associations within the HLA system.

Notes: Having reviewed the status of H-Y as the sex-determining antigen concerned with the differentiation of teh dominant gonad, we consider some of the problems deriving from the tests for this antigen, and from their application to the study of natural experiments. To reconcile the results of these studies with the alleged influence of H-Y on gonadal development, we propose and discuss a hypothesis on the genetic control of the synthesis of this antigen. This states that an autosomally-coded, positively cross-reacting precursor is rendered biologically active by a Y-chromosomal gene, and transformed (in a dosedependent manner) into a biologically inactive, antigenically negative substance under the influence of an X-chromosomal gene.

Notes: The presence of H-2Ld antigens was evaluated in methylcholanthrene-induced BALB/c fibrosarcomas by a varity of approaches. Transplantation experiments showed that BALB/c-H-2dm2 mice, a mutant strain whose cells do not express H-2Ld antigens, after immunization with BALB/c normal tissues developed a resistance to the growth of two tumours (C-3 and GI-17), but not to a third neoplasm, C-1, which is known to have H-2d-as well as H-2k-like alien antigens. In vitro experiments with cytotoxic T lymphocytes generated against Ld antigens confirmed a loss of Ld antigens on C-1 but not on C-3 tumour cells. Serological experiments with an anti-Ld serum again revealed the presence of H-2Ld determinants on C-3 but not on C-1 cells. Biochemical analysis in SDS-PAGE of immunoprecipitates obtained by specific anti-H-2 sera with NP40 lysates of the tumours studied could detect H-2Kd, H-2Dd and H-2Ld antigens in C-3 fibrosarcoma cells whereas Kd and Dd were the only H-2d molecules found in C-1 lysate along with the H-2k-like specificities. The possible genetic mechanisms which may explain this apparent gain and loss modification of the H-2 profile of C-1 are discussed.

Notes: Recently, in investigating the responses of T-cell from high responder mice that were primed with myoglobin (Mb) or with synthetic peptides containing antigenic site 5 and increasing in length stepwise by increments of two residues, we observed that T-cell recognition was highly dependent on conformation. In the present studies, tolerization experiments were carried out to further investigate this finding. Neonatal mice (BALB/cByJ) were either tolerized with Mb or with synthetic peptides of Mb containing antigenic site 5. Tolerization with Mb and subsequent immunization with Mb gave T-cells that did not proliferate in vitro to Mb or any of the peptides. T-cells from mice that were tolerized with a truncated peptide 139-153 (having deletions at Tyr-151 and Ala-144) and subsequently immunized with Mb proliferated in vitro to Mb and to peptides 132-153, 135-153 and 143-153. T-cells from mice that were tolerized with native Mb and subsequently immunized with peptides (which are unfolded in solution) did not proliferate in vitro to Mb, but responded well to the peptides. Conversely, tolerization with peptides had no effect on the recognition of, and the response to, native Mb by the T-cells, whereas the response to the peptides was completely removed. It was thus concluded that the recognition of protein antigens (or at least of Mb) by T-cells is (like the recognition by antibody) dependent on the conformation of the antigen.

Notes: Gm phenotype frequencies were examined in 112 Swedish myasthenia gravis patients. The G1m 1,2,3 phenotype frequency in the total patient material did not differ significantly from that found in the normal population. However, when patients were subdivided, three different patient groups were observed with regard to Gml frequency: (1) Thymoma patients having a low frequenc of Gm1, (2) Non-thymoma patients with a mild disease having a low frequency of Gm1 and (3) Non-thymoma patients with a severe disease having a high frequency of Gm1. When patients were subdivided according to presence or absence of HLA-B8 and Gm1 respectively, severe symptoms were less frequent in the HLA-B8+, Gm(-1) group as compared to the HLA-B8+, Gm(+1) group. Furthermore, there was an increased frequency of sera with anti-immunoglobulins not inhibitable by pooled control immunoglobulins.

Notes: In the preceding communication of this series, the fine specificity requirements for T-cell recognition of one of the Mb antigenic sites (antigenic site 5) were examined. Seven synthetic peptides containing antigenic site 5 and progressively increasing in length to the left by increments of two residues up to 22 residues in length were studied with regard to their ability to stimulate T-cell proliferation of Mb-primed T-cells from high responder H-2d and H-2s mouse strains. Unexpectedly, it was found that, unlike smaller and longer peptides, some intermediate-sized peptides failed to stimulate T-cell proliferation even though they contained the full antigenic site. This indicated that lack of proliferative stimulating activity by a peptide does not necessarily imply absence of an antigenic site in the peptide. The results enabled us to conclude that T-cell recognition of native proteins (or at least of Mb) is dependent on protein conformation. In the present work, we have examined the ability of peptide-primed T-cells from H-2d and H-2s mouse strains to proliferate to Mb. Immunization with peptide 145-151 (antigenic site 5) afforded T-cells that did not proliferate in vitro to Mb or any of the synthetic peptides 145-153, 143-153, 141-153, 139-153, 137-153, 135-153 or 132-153. When mice were primed with either peptide 143-153 (11 residues) or peptide 132-153 (22 residues), The T-cells obtained did not respond to Mb but responded in each case very well to peptides 132-153, 135-153, 137-153 and gave lower, but significant, response to the shorter peptides 145-153 and 143-153. Intermediate-sized peptides did not stimulate T-cell proliferations and neither did truncated peptides that had deletions at Tyr-151 and Ala-144. These findings underscore the fine specificity of the T-cell and indicate a high level of conformational dependency for T-cell recognition. It is also concluded that macrophage recognition and presentation of protein antigen must involve the intact native protein and it is probably not related to the processing and fragmenting of the antigen by macrophage. The latter activity is more related to the role of macrophage in clearance.

Notes: All classes of molecule postulated to play a role in recognition or to have some structural homology to immunoglobulins were compared by the SΔQ index of differences in amino acid composition. The results suggest that there is a wide family of immunoglobulin-related molecules that includes vertebrate immunoglobulins, T cell receptors, MHC products, C-reactive proteins, and (some) invertebrate lectins.

Notes: IgM and IgG mouse monoclonal antibodies with specificity for the blood group B determinant have been produced by immunization of mice with a partially purified salivary glycoprotein. These antibodies have been characterized and one of the IgM antibodies showed potential as a grouping reagent. The ready availability of these salivary blood group substances offers the potential to produce a wide range of related monoclonal antibodies.

Notes: No evidence was found for close linkage between the human C6 gene (6th component of complement) and 28 marker loci, using the LIPED 3 computer program of Ott (1974). But the data allow the exclusion of the C6 locus from large parts of the autosomes 1, 2, 4, 6, 8, 9, 13, 14, 16, 19, and 20.

Notes: The humoral response of twelve strains of mice to immunization with either normal adult human haemoglobin (HbA1) or sickle-cell haemoglobin (HbS) was studied using a quantitative enzyme-linked immunosorbant assay (ELISA). Results indicate the presence of at least two genes that control the magnitude of this response, one of which is located at the ‘D-end’ of the H-2 complex. H-2 haplotypes s, q and k are associated with a low response, haplotypes d and a with a high response, and the b haplotype with an intermediate response. No difference in the pattern or magnitude of the response was observed when HbS was used as the immunogen in place of HbA1. The N-terminal octapeptides of the β chains of HbA1 and HbS were synthesized and the presence of antibodies specific for the N-terminal octapeptides was tested. The magnitude of the anti-N-terminal response correlated with that to the intact molecules with one exception. A much lower response to the βs-peptide was found in the C57BL/10Sn mice. Monoclonal antibodies were prepared to HbA1 and HbS. Three clones were studied for their reaction with HbA1' HbS, and the βA1 and βs peptides. One clone was found which binds both the βA1 and βs peptides. This indicates that the N-terminal portion of the β chain of human haemoglobin is antigenic in mice.

Notes: Erythrocytes that exhibit the rare blood group p phenotype lack the P antigen (globotetraosylceramide) and the Pk antigen (globotriaosylceramide). This phenotype is inherited as an autosomal recessive condition and the red cells of heterozygous individuals, parents and children of p persons, are serologically normal but no chemical analyses of their red cells have been reported. We have studied an unusual family in which all five children exhibit the p phenotype. In addition to the abnormalities described previously, the erythrocytes of four siblings had twice the normal concentration of lactotriaoslyceramide and lactoneotetraosylceramide. These cells also contained 3-5 times as much sialosyllactoneotetraosylceramide and up to a two-fold increase in GM3 ganglioside. The glycolipids of the parents' erythrocytes were normal. Electrophoretic analysis of the glycoproteins of the proposita's erythrocytes revealed no abnormalities, but her erythrocyte membranes contained approximately 35% less galactosamine than normal red cells. This abnormality resulted from a marked decrease in galactosamine that was soluble in chloroformmethanol. The lipid-extracted residue, which contained the glycoproteins, had a normal galactosamine content.

Notes: The identity and complete purification of mouse Thymocyte Interaction Modulation Factor (T IMF) is described. Use of silver-stained PAGE methods shows that previous methods of purification yield preparations containing two protein or glycoprotein bands. T IMF activity from H-2k mice can be bound to 15.5.5 monoclonal antibody columns (anti H-2 Dk) but not to 11.4.1 columns (anti H-2 Kk). The activity can be recovered from 15.5.5 columns and runs on PAGE as a single band at approximately 34,000 Daltons. This evidence together with previous evidence relating the activity to H-2 D locus argues that T IMF is a soluble H-2 D antigen fragment equivalent to a papainized H-2 fragment. Additional evidence is presented on an improved method of assay of T IMF activity, on its inactivation by enzymes and serine-esterase inhibitors and of its effect on syngeneic leucocytes and macrophages. It is also shown that T IMF is not appreciably toxic for cells.

Notes: One hundred and sixty-five individuals with ragweed rhinitis and asthma were studied. The associations between the HLA system and: Ra3 skin-test response, Ra3 RAST, Ra3/antigen E skin-test reactivity ratio, Ra5 skin-test response, Ra5 RAST, Ra5/antigen E skin-test reactivity ratio, serum IgE levels, antigen E skin-test response, and antigen E RAST were studied. In addition, the influence of the serum IgE levels of these immune parameters were evaluated. No highly significant associations were noted.

Notes: We have previously shown that the antibody response to mammalian myoglobins is under genetic control. In the present study we examined antibodies to sperm-whale, Atlantic bottlenosed dolphin, Black Sea dolphin, horse and badger myoglobins, raised in high responder strains of mice, to ascertain whether there is genetic control of antibody affinity to mammalian myoglobins. Using antisera of varying dilutions, the binding to 125I-labelled homologous myoglobins was studied by inhibition with homologous myoglobin over a wide range of inhibitor concentration in a modified Farr assay. The results indicated that there are no large differences between high responder strains of mice in the affinity of antibodies to mammalian myoglobins.

Notes: No. Gm, Am, Pi, and Km gene frequency difference has been observed between eighty-seven Caucasin IgA nephropathies and ninety-six controls. But a Km1 significant increase is found in patients with chronic renal failure compared with the total patients group. This data fits for a genetic predisposition factor in IgA nephropathy.

Notes: The genetic control of the in vivo growth of the Moloney virus-induced BALB/c lymphoma YC8 was studied in F1 hybrids between BALB/c and several strains differing at the MHC and/or at the level of non-H-2 genes. Parental strains of the B10 and C3Hf but not of A, BALB/c or DBA/2 backgrounds introduced a significant resistance to the growth of 102 YC8 cells (a dose able to kill 100% of BALB/c mice) in semisyngeneic hosts. This resistance appeared to be due to non-H-2 genes although a modulation of the tumour growth by genes encoded by the MHC was also evident. The study of backcrosses between susceptible BALB/c and resistant (BALB/c x B10.BR)F1 crosses revealed that 83% of animals developed lethal tumors after injection of 102 YC8 cells. This high frequency of tumour takes was not linked to genes of the MHC. Adult thymectomy plus sublethal irradiation was able to abrogate the resistance of (BALB/c x B10.BR) or (BALB/c x B10.RIII)F1 mice to YC8 growth. Since the injection of silica also impaired the resistance to YC8, we tentatively concluded that the genetic control of resistance to YC8 is mediated both by T cells and macrophage-like cells.

Notes: Mice of independent haplotypes and several recombinant inbred strains were immunized with highly purified preparations of either the α-chain or β-chain subunit of human adult haemoglobin. Cells from the sensitized lymph nodes were challenged in vitro with the appropriate subunit (or in some cases both chains) and cell proliferation assessed by 3H-thymidine incorporation. Mice of the H-2b and H-2d haplotypes were high responders to α-chain while mice of the H-2f, H-2j, H-2k, H-2r, H-2s, H-2u, and H-2v haplotypes were low responders. The low responsiveness of B10.A(4R) and B10.MBR and the high responsiveness of B10 indicated that the Ir gene(s) determining responsiveness to the α-chain subunit resides in the I-A subregion of the mouse major histocompatibility complex. Mice of the H-2d, H-2f, and H-2s haplotypes were high respoders and H-2b, H-2f, H-2q, and H-2u haplotype mice were low responders to β-chain. H-2k, H-2p, H-2r, and H-2v haplotype mice were intermediated responders to β-chain. The low responsiveness of B10.S(8R) and B10.TL and the high responsivenes of B10.S(9R) mapped the Ir gene(s) to β-chain to the I-A subregion. Data collected from challenging high responder cells with both subunits indicated that α-chain and β-chain do not crossreact. These results are discussed in reference to earlier observations suggesting that the low responsiveness of some strains of mice to priming and challenging using the intact haemoglobin molecule might be due to a negative regulatory influence mediated by one of the subunits. In the absence of this influence these mice would respond normally.

Notes: An attempt to determine association between immunoglobulin allotypes and Schistosoma japonica infection was made. The results show that no apparent relationship exists.

Notes: Diet and Resistance to Disease; Advances in Experimental Medicine and Biology. Vol. 135. M. Phillips and A. Baetz

Notes: The antigenic specificities of six (1-6) IgG allotypes of the domestic mink were tested in the sera of closely related species of Mustelidae family and distant mammalian species. It was found that allotypes 1 and 5 are ancient. Their antigenic specificities were established not only in Mustelidae, but also in other taxonomic orders of mammals. Allotypes 3 and 2 are phylogenetically younger; they were detected only in Mustelidae. Allotypes 4 and 6 appear to be unique to the domestic mink.The instantaneous evolutionary emergence of complex allotypes 4 and 6 is difficult to explain by a rapid accumulation of gene point mutations during phylogenesis. Activation in the domestic mink of those immunoglobulin genes, which are silent or poorly expressed in closely related Mustelidae, is suggested as a more plausible explanation.

Notes: In the V/V type of bovine erythrocytes (BE), the antigen IM and the Thomsen-Friedenreich (TF) receptors, which reacted with a TF-specific agglutinin from human sera and the lectin from Helix pomatia, were all destroyed by treatment with trypsin. Unlike the V/V, in the F/F type of BE the antigen IM and these TF receptors were not only not destroyed by trypsin, but this enzyme treatment was, in fact, necessary to make them amenable to the corresponding agglutinins.Another TF receptor, which reacted with the lectin from peanuts, was identified on a trypsin-resistant component of all BE and independently of the FV genotypes.

Notes: Five major glycoproteins with anti-B agglutinin activity were isolated from seeds of Euonymus Sieboldiana by a procedure based on precipitation with ammonium sulphate, Sepharose 4B gel filtration, CM- and DEAE-Sepharose chromatography and Sephacryl S-200 gel filtration. The purified glycoproteins each gave a single symmetrical peak on Sephacryl S-200 gel filtration with elution volumes corresponding to molecular weights of approximately 15,000 to 130,000, each forming a single precipitin line on gel diffusion plates with anti-E. Sieboldiana antibody. These anti-B glycoproteins were rich in acidic amino acids without cysteine and methionine and contained about 8-36% carbohydrate, of which galactose, arabinose and glucose were the predominant sugars, with small amounts of glucosamine, rhamnose, fucose, xylose, mannose and ribose. The most anti-B active lectin agglutinated human B red blood cells at a concentration of 2 μg/ml and was strongly inhibited by melibiose. The three other lectins with anti-B agglutinin activity, however, were not inhibited by any galactose-containing glycosides.

Notes: Linkage between the Pi (alpha1-antitrypsin) and Gm (immunoglobulin heavy chain) loci was studied in thirty-four families including forty-one informative parents and 142 children. In females, the results did not provide evidence for linkage (posterior probability of non-linkage 0.98). In contrast, in males, there was strong evidence for linkage (peak lod 3.9 at 0= 0.18, posterior probability of linkage 0.98). The two populations appeared to be significantly different (0.001 〈 P 〈 0.01) with respect to the heterogeneity criterion of Morton. In addition, the effect of the possession of the S allele (associated with significantly decreased serum alpha1-antitrypsin levels) was studied in fifteen informative parents and fifty-three children of the same group. No evidence for or against linkage was found in females, but in males close linkage between Pi S and Gm was demonstrated (peak lod 7.7 at 0= 0.05, posterior probability of linkage 0.9999). These data indicate significant linkage between Pi and Gm in males but not females and close linkage between the Pi S and Gm markers in males.

Notes: Cytotoxic antibodies to HLA-A,B,C and to Ia-like antigens were detected in about 8% of sera drawn from 1312 women several years after their last pregnancy. In the majority of sera anti Ia-like antigen antibodies were not associated with anti HLA-A,B,C antibodies. Persistence of both types of antibodies was not correlated with the number of pregnancies and with the time interval between the last immunizing stimulus and the drawing of the sample. Testing of the sera with a panel of HLA typed lymphocytes identified 20 sera specific for HLA-A,B alloantigens and 7 specific for Ia-like alloantigens.

Notes: The evidence of the past 10 years indicates that genes mapping in the I region of the mouse major histocompatibility complex (H-2) regulate a bewildering array of immunologic functions. A survey of H-2-linked specific immune response (Ir) genes shows that the phenotypic effect of these genes is to enable a particular inbred strain to discriminate and recognize remarkably precise chemical specificities, such as one or two amino acid interchanges in a polypeptide chain. The only I region gene products which have been identified to date are the Ia antigens. These include five readily detectable polypeptide chains (Aa, Ab, Ae, Eα, and Ii) and several other serologically distinct entities which are selectively expressed on functionally distinct T cell subsets (J1, J2?, J3? and C). The specificity of recognition of Ir genes would seem to require a larger number of I region gene products than can be generated even by combinatorial association of the four readily identifiable peptides (to give eight combinatorial associations) and the other serologically identified gene products. If the Ia antigens are to function as an antigen specific receptor system, separate from immunoglobulin molecules, there must be other, as yet undetected, I region gene products (e.g. Ia antigens). Alternatively, the known I region gene products could function by any one of several postulated mechanisms to generate an antigen specific T cell receptor system. The available evidence for the total number of I region gene products is reviewed, and the alternate possibilities are briefly discussed in this presentation.

Notes: Different patterns of tumour growth resulted from inoculation of separate isolates of the Subgroup C Bratislava 77 strain (B77) of Avian Sarcoma Virus (ASV) into three closely-related inbred lines of chickens. The major genetic difference between these chicken lines is that each is homozygous for a different MHC haplotype. Since for one of the viral isolates, resistance to progressive tumour growth has been shown to be controlled by MHC-linked genes, the data presented here suggest that MHC-controlled tumour rejection operates on viral or cellular determinants different from those which define classical viral group or subgroup specificity.

Notes: The red cells of a normal male blood donor, K.S., were first grouped as B but he was found to lack anti-A in his serum. Closer investigation revealed that his red cells had very weak A activity, demonstrable only by absorption and elution of anti-A. He is a non-secretor of ABH and a secretor of Lea. Blood group A-, B and H-gene specified glycosyltransferases were detected in his serum. In contrast to the finding of a B antigen of normal strength on his red cells, the B transferase in his serum was only about 30% of the normal level and, despite the very weak A activity of K.S.'s red cells, the A transferase level was about 50% of that found in the serum of group A individuals with normal strength of A antigen. Moreover, the A transferase on the basis of its pH optimum, Km values for donor and acceptor substrates, activation by divalent cations, isoelectric focusing profile and capacity to convert O to A-active cells, was characterized as the product of an A1 gene. A family study showed that K.S.'s wife is group A2 and that they have two sons, one group A2 and the other group B. The group B son is assumed to have inherited a B gene from the propositus but the level of B transferase in the son's serum is three times as high as that in his father's serum. The wife of the propositus and his group A2 son have normal A2 transferases in keeping with their A2 red cell status. The A2 son therefore appears to have inherited an A2 gene from his mother but neither the A1 nor the B gene shown to be carried by his father. The distribution of transferase activities in K.S.'s red cells differs from that in his serum. A level of B transferase within the normal range was found in his red cell membranes but a very low level of A transferase was detected. The discrepancies between the serum transferases and ABO-red cell group, together with the pattern of inheritance within the family, led to a suspicion of chimaerism. This was confirmed by the finding of fibroblasts with the female 46XX karyotype in cultures of the propositus' skin. These results suggest that K.S. is a dispermic chimaera with two different cell lines of the genotypes BO and A1O or A1A1. The group A2 son is assumed to have inherited an O gene from his father. It seems probable that K.S.'s bone marrow and reproductive organs are comprised predominantly of the XY cell line which carries the blood group BO genotype whereas his skin and other tissues which contribute the A1 transferase to his plasma, are partly made up of the XX cell line which carries the blood group A1O or A1A1 genotype.

Notes: A cloned hybridoma, producing an IgM kappa antibody, has been derived from a fusion between NSI myeloma cells and spleen cells from BALB/c mice immune to CBA lymphocytes. Its specificity for Lyt-1.1 alloantigen has been established from (1) the lymphoid tissue distribution restricted to T cells and (2) the reactivity pattern on a panel of thymocytes from standard and Lyt congenic mouse strains.

Notes: Haemagglutination inhibition studies using isolated membrane components from rat red blood cells showed that the activity of the major antigenic system RT2 was contained exclusively in the glycoprotein fraction and confirmed immunochemically the existence of two alleles, RT2a and RT2b. The SDS-PAGE analysis of the membrane showed two interconvertible glycoproteins of apparent molecular weight 72,000 and 36,000, and the higher molecular weight component appeared to be a dimer of the lower molecular weight one. Analysis of the various membrane preparations after 125I-labelling showed that the glycoproteins existed in at least two molecular species on the surface of the red cells and that one species was more readily radio-iodinated and dimerized than the other. The RT2 antigenic specificities were preferentially associated with this component.

Notes: Spleen cells from C3HfB/HeN mice (H-2kv1) were markedly less susceptible to lysis by monoclonal anti-Kk antibody (Clone 11-4) than spleen cells from C3H/HeN mice (H-2k). Less than 50% of C3HfB/HeN cells were lysed even at the highest antibody concentration tested compared to 70% of C3H/HeN cells. The difference in end point titre of the antibody tested on cells from the two strains was 5-10-fold. Similar differences were not observed using the conventional H-2Kk-typing antiserum (D-23).

Notes: F. H. BACH, B. BONAVIDA, ELLEN VITETTA, AND C. F. Fox; T and B lymphocytes: recognition and function. Academic Press, New York, 1979. ISBN 0-12-069850-1, Price U.S. $36.50.

Notes: The RTI-B/D region-associated antigens which were serologically defined in the previous study (Ohhashi et al., 1981), were partially purified from membranes of a rat B cell leukaemia, KNL-14. Sequential immunoprecipitation test, with the partially purified 125I-B/Dak preparation using four different rat alloantisera, including a monoclonal antibody, disclosed three distinctive populations of β units of the class II molecules. Highly purified β units of three discriminable class II molecules were shown to have different structural properties in terms of molecular weights and of electrophoretic profiles on the isoelectric focusing. The β units shifted to a position of higher molecular weight on SDS-PAGE under reducing condition, thus suggesting to carry intradisulfide bonds. Furthermore, the highly purified β units crossreacted with murine anti-Ia sera. The rebinding test revealed that at least two discriminable species of β units cross-react with anti-I-Ak monoclonal antibody, whereas β units purified by binding with the IE4 monoclonal antibody cross-reacted with anti-I-Ab and/or anti-I-Ad antiserum. On the basis of structural and antigenic properties, we have postulated that the rat class II region can be divided into at least three subregions, each containing a locus which encodes a distinctive β unit of the class II molecule.

Notes: In attempts to identify cell surface molecules specified by lethal genes in the T/t-complex, we prepared a rabbit antiserum that has cytotoxic activity against testicular cells from males heterozygous for t12, but not against wild type cells. However, anti-t12 serum immunoprecipitates the same major component, a glycoprotein of mol. wt. 87,000 daltons, from galactose-labelled C3H.+/t12 testicular cell lysate and from congenic C3H.+/+ lysate, although the gp87 molecule precipitated from +/t12 cells appears to be more highly galactosylated than the +/+ form. The antigen is heavily glycosylated in both genotypes, since when testicular cells are treated with tunicamycin before immunoprecipitation, a protein of 40,000-42,000 daltons is obtained.Gp87 is also present on pre-implantation embryos, and on teratocarcinoma cells, but is barely detectable on any adult somatic cells examined. Its expression is developmentally regulated during pre-implantation stages, but the temporal pattern of its expression appears to be different between wild type and t12 embryos. Thus, we believe we have identified a molecule that may play a role in the differentiation of testicular cells and pre-implantation embryos, and that is either specified by genes in the t12 haplotype, or responsive in some way to the effects of t12.

Notes: Five monoclonal antibodies reacting with class I MHC antigens were produced by fusing lymphocytes from WF (RT1u) rats immunized against DA (RT1a) rats with P3-X63-Ag8-653 myeloma cells. Sequential immunoprecipitation studies with the mAb and the WF anti-DA alloantiserum demonstrated the presence of four different class I molecules: all four molecules were reactive with the alloantiserum; three of them contained the determinant for mAb 155; two of the latter three molecules shared the determinants for mAb 3, 56 and 60, and one of these two molecules also contained the determinant for mAb 118. The four molecules could be isolated from the antigen preparation by sequential immunodepletion first with 118, next with 3, then with 155 and finally with the alloantiserum or by sequential absorption with affinity columns of Sepharose 4B coupled to the antibodies. The three antigens which were sequentially isolated with the mAb 118, 3, and 155, respectively, were analysed by SDS-PAGE after digestion with Staphylococcus aureus V8 protease, and they showed differences in peptide patterns. The relative amounts of the antigens expressed on red blood cells and on lymphocytes were different based on the results of sequential isolation and indirect cellular radioimmunoassay: the antigen which reacted with both mAb 3 (and 56 or 60) and 155 was the major class I antigen on red blood cells, and the antigen which reacted with mAb 118, 3 (and 56 or 60) and 155 was the major class I antigen on lymphocytes.

Notes: The frequency of dorso-ventral vaginal septa (DVS) in mice in influenced by genes associated with the major histocompatibility complex, H-2. Data shows that two subregions within the H-2 complex, K-Eβ and D, have a very strong influence on the frequency, while the Eβ-S subregion appears to have a weak effect.

Notes: Two directional selections of rats for a high or a low IgM serum levels have been carried out. The criterium of selection was the individual merit of each rat in having a high or low IgM level when they were 3 months old. All animals were kept together, in order to avoid external influence on the IgM synthesis. The founding population was a mixed nucleus of rats, obtained by breeding of 13 different strains of inbred or outbred rats for two generations. Two lines, one with a high IgM serum level and another with a low IgM serum level, have been separated. At generation 10, they differed from each other by a coefficient of approximately four. These IgM serum levels were not due to differences in IgM catabolism of the two lines since both lines had similar IgM half-lives. Analyses of the data obtained show that, for IgM synthesis, the coefficient of heritability varies between 0.30 to 0.40, and the number of `independent loci' controlling IgM synthesis ranges from 11 to 14.

Notes: Two murine monoclonal antibodies (E11-1 and MR4-130) agglutinated all samples of human red cells except those of the Ge (-1, -2, -3) phenotype. It was possible to demonstrate that these antibodies recognize two different epitopes of the Gerbich antigen.

Notes: Genetic polymorphism of human C6 was investigated in Japanese using isoelectric focusing and a specific haemolytic overlay method. Three common and six rare allotypes were identified. Five of these nine allotypes were reference-typed by the International Reference Laboratory. Five of the six rare allotypes were considered to be new. The allele frequencies were estimated in the population study as follows: C6 A 0.427, C6 B 0.483 C6 B2 0.076, and the rere alleles (A3, A21, M1, M2, B3, and B4) 0.041.Inheritance of the three common and the two rare (A3 and M1) allotypes was demonstrated in the family study. The patterns obtained by the pretreatment with neuraminidase are presented.

Notes: Previous studies in this laboratory have resulted in the determination of the antigenic structure of myoglobin. The present work was carried out to investigate the genetic control of the murine antibody response to myoglobin following immunization with free (i.e., not coupled to a carrier) synthetic antigenic sites or other peptides corresponding to surface regions of myoglobin that are not immunogenic when the native molecule is the immunizing antigen. Synthetic peptides corresponding to antigenic site 1 (peptide 15-22), site 2 (peptide 56-62), site 3 (peptide 94-100), site 4 (peptide 113-120), site 5 (peptide 145-151) and two surface regions, peptide 1-6 and peptide 121-127, were injected in complete Freund's adjuvant in different strains of mice. Serum antibodies specific for myoglobin were subsequently obtained and were measured by means of a radioimmune plate binding assay in which Mb was used as the solid phase antigen. It was found that the genetic control of the antibody response to myoglobin following immunization with the free synthetic peptides was different from the genetic control obtained following immunization with native myoglobin. The significance of this finding is discussed.

Notes: A β-galactoside α1 → 2 fucosyltransferase (H-enzyme) from human group O plasma which provides H blood group specificity to erythrocyte membranes has been purified approximately 22,000-fold by chromatography on DEAE-Sepharose CL-6B, GDP-hexanolamine-Sepharose 4B and SP-Sephadex C-50. The molecular weight of the H-enzyme was estimated to be 150,000 by gel filtration. Human group O erythrocyte membranes which had lost their H blood group activity by the action of α1 → 2 fucosidase were fucosylated by the transferase, and restored the H activity. Radioactive L-fucose appeared to be incorporated into glycolipid blood group substances of erythrocyte membranes. The activity of the α1 → 2 fucosyltransferase from human plasma, stomach mucosa, erythrocyte membranes and porcine stomach mucosa were specifically inhibited by the rabbit antiserum immunized with the preparation of human plasma H-enzyme. The anti-plasma H-enzyme antiserum did not inhibit the activities of α1 → 3 N-acetygalactosaminyltransferase (A-enzyme), α1 → 3 galactosyltransferase (B-enzyme), and β-N-acetylglucosaminide α1 → 3 and α1 → 4 fucosyltransferases from human plasma and stomach mucosa.

Notes: The monoclonal antibody H8, previously described as anti-JMH, has the same specificity as a JMH-related antibody, R.M. H8 blocks the reaction of human anti-JMH and related antibodies with JMH+ cells, suggesting that the JMH-related antigens are very closely situated to each other on the red cell membrane.

Notes: The existence of HLA-encoded genetic factors controlling susceptibility to tuberculoid leprosy in humans has been firmly established. Furthermore HLA-DR2 has been recognized as a marker for tuberculoid leprosy in India. At this moment, however, the gene products involved and the mechanism by which they confer susceptibility to tuberculoid leprosy remain only speculative. In an attempt to detect in vitro the expression of these HLA-encoded factors, we studied 12 tuberculoid leprosy patients and 22 healthy family members in a lymphocyte transformation test (LTT). All individuals were derived from multi-case Indian families, previously reported to show the presence of HLA-linked susceptibility genes. Although the responder-status of the healthy contact siblings was shown to behave independently from the fact whether they were HLA-identical with the patient-siblings or not, some evidence for in vitro expression of HLA-DR2 associated factors could be obtained. Nevertheless, it is concluded that the standard LTT seems not to be a test-system of first choice to detect the in vitro expression of the genes under study.

Notes: The survival time of skin allografts was investigated in 28 combinations of mouse strains differing at various loci, in adult recipients treated with lentil seed lectin (LCA). The recipients were given 1 mg of LCA daily (i.v. or i.p.) after transplantation of the skin graft. The immunosuppressive effect of LCA was generally inversely related to the conventional strength of the allotransplantation reaction, i.e. the rate of rejection of allografts on untreated recipients. In the case of H-2D antigenic disparities, the grafts usually survived for more than 100 days. The H-2A or H-2K disparity appeared to be relatively resistant to induction of tolerance by LCA treatment. The relative resistance of various H-2 antigens toward the induction of tolerance by LCA differs to some extent from that observed in experiments on the induction of neonatal tolerance by semiallogeneic lymphocytes as described in literature.

Notes: In mice it has been shown that mating preference is genetically associated with the Major Histocompatibility Complex (MHC). In the present study, an approach is described to study homologous aspects of the MHC in man.After selection of families with one or more children, a given parental HLA antigen was selected and the frequencies of the spouse's HLA antigens were determined. Assuming a random distribution, these frequencies should not be statistically different from those of the total population.Evidence is presented that this distribution is not random for a number of maternal and paternal HLA antigens.

Notes: We compared H-2D products of B10.D2 and the mutant B10.D2-H-2dml spleen cells using two monoclonal antibodies designated D3.179 and S11.7. D3.179 reacts with public specificities and appears to define more than one product as shown by cytotoxicity and binding studies. Strong reactivity was obtained in both binding and cytotoxicity and binding studies. Strong reactivity was obtained in both binding and cytotoxicity to B10.D2 and marginal cytotoxicity and low binding were observed with B10.D2-H-2dml cells. The second monoclonal antibody S11.7, in contrast, reacts only with H-2Dd bearing strains and gave strong binding to B10.D2-H-2dml and weaker binding to B10.D2. Taken together, the observations indicate that the mutant not only expresses H-2D, encoded antigens but bear quantitatively greater amounts of some specificities than the parent. A Product of approximately 46,000 daltons was immunoprecipitated from B10.D2 cells by D3.179. Very small amounts of this product were seen in B10.D2-H-2dml immunoprecipitates.

Notes: Abstract. Five species of fingerling fish, channel catfish, Ictalurus punctatus, tilapia, Sarotherodon aureus, golden shiner, Notemigonus crysoleucas, largemouth bass, Micropterus salmoides and bighead carp, Aristichthys nobilis, were tested to determine their susceptibility to the bacterium, Edwardsiella ictaluri, at 26°C, Channel catfish demonstrated high susceptibility to E. ictaluri as 100% of those fish injected with 1.5 × 103 cells died within 10 days. Tilapia demonstrated slight susceptibility to the pathogen while golden shiner, bighead carp and largemouth bass were not susceptible. E. ictaluri was isolated from a higher percentage of peritoneal cavities, livers and kidneys of channel catfish than of other species. Sequential growth of E. ictaluri in the liver of channel catfish is described.

Notes: Abstract. Eimeria branchiphila sp. nov. is described from the roach, Rutilus rutilus L., collected in Bulgaria. Sporulated oocysts were found in gill secondary lamellae and less frequently were associated with melano-macrophage centres in the spleen and in kidney interstitial tissue. In the gills, parasite cells identified as zygotes and sporulation stages were also encountered. This unusual site of sporulation represents a mechanism of sporocyst dispersal previously not recorded in fish coccidia.

Notes: Abstract. Fifteen drugs were tested for in vitro control of Yersinia ruckeri. Those drugs yielding in vitro control were subsequently tested for in vivo control of induced enteric redmouth disease in rainbow trout. Two of the drugs, Tribrissen®v and tiamulin, controlled Y. nickeri in vivo under laboratory conditions. The effective doses used in this study were 1–0 mg of Tribrissen/kg fish/day and 5–0 mg of tiamulin/kg fish/day, when the compounds were administered orally for 14 days. No toxic effects were noted at these dose levels, or from Tribrissen given at 15 times the above effective dose.

Notes: Abstract. S, a dominant, lethal, atitosomal gene, was discovered in Auburn University's population of Surotherodon aureus (=Tilapia auera). When present in the heterozygous state (S+). the S gene produces the saddleback phenotype. Saddlebacks are missing part of or all of the dorsal fin. Some fish thai are missing most of or the entire dorsal fin have additional fin disorders and are missing various combinations of the pectoral, pelvic, and anal fins. Each fin disorder is associated with gross skeletal anomalies. Viability in saddlebacks is reduced by 67% during the first three months of life. Those that survive are less resistant to stress and subsequent infection than normal fish. When challenged by Saprolcegnia sp., saddlebacks were far more susceptible to infection (P〈0.01). When present in the homozygous state (SS), the gene causes death in fry prior to swim-up. Thus, a single S gene reduces viability by 67%, and two S genes reduce viability by 100%. The S gene did not affect early growth or dressing percentage. Because it is a dominant gene, the S gene can be eliminated from any population in one generation by culling all saddlebacks.

Notes: Abstract. The histopathological changes in the eye due to the penetration and growth of the parasite, Lernaea piscinae, in the host tissue of big head carp is described. Distinct pathological changes were hyperplasia of the corneal epithelium, severe inflammatory response mainly around the anchor of the parasite, in the anterior chamber, the corneal stroma and the vitreous body and detachment of the vitreous and retinal layers. Rupture of the lens capsule and fibrous tissue response around the process of the anchor structure of the parasite were also observed. The development of an immune response to the parasite within the body of the host is thought to have led to the infection of the cornea, an avascular site.

Notes: Abstract. Steelhead trout were exposed to sublethal concentrations of copper and inoculated with Yersinia ruckeri. Copper exposure at 7 and 10 μg/1 for 96 h caused more fish to die of infection than control fish (no copper). Infection susceptibility increased with time of exposure to a single dosage of copper (10 μg/1), reaching a maximum at 48 h. Lowering the copper concentration to 5 μg/1 caused the infection susceptibility to occur at 24 h. The infectious dose of Y. ruckeri was lower in fish exposed to 10 μg/1 copper for 48 h than control fish.

Notes: Abstract. Four species of flatfish [plaice, Pleuronectes platessa L.; turbot, Scophthalmus maximus (L.); Dover sole, Solea solea (L.) and the common dab, Limanda limanda (L.)] were experimentally infected with cercariae of Stephanochasmus baccatus under similar conditions and examined at intervals up to 7 weeks post-infection- The tissue response was found to be qualitatively the same in all four hosts and was consistent with a chronic granulomatous inflammatory reaction induced by resistant particulate foreign bodies. The intensity and rate of the response varied between host species but the capsule formation was essentially similar in all. The mature capsule consisted of an inner layer composed of leucocytes which underwent epithelioid transformation. The cellular structure of this tissue gradually degenerated from the oldest layer to form a dense eosinophilic ground substance in which was embedded nuclear debris. The outer layer of the capsule consisted of fibroblasts and reticulin collagen fibres. Muscle and fin cysts differed, with muscle cysts tending to be less discrete and more fibrous. The rate and intensity of the response to the parasite varied between different host species. Plaice and turbot showed the most intensive response and in turbot the reaction was considerably accelerated. Mortality of encysted metacercariae was only seen in turbot. The overall effect on larger 1-group fish was minimal, but the smaller 0-group plaice were more severely affected by the relatively large cvsts that interfered with organ function.

Notes: Books reviewed in this article:Fischkrankheiten 4th edn. By W. Schäperclaus. Akademie Verlag, Berlin, 1979, 2 volumes, 1089 pp.Ecology of Freshwater Fish Production. Ed. by Shelby D.Gerking. Black well Scientific Publications, 1978, 520 pp.Diseases of Fishes. Book 6. Fungal Diseases of Fishes. By G. A. Neish & G. C. Hughes. TFH Publications, New Jersey, 1979, 159 pp.Developments in Deep-Sea Biology. By N. B. Marshall. Blandford Press, Poole, Dorset, U.K., 1978. 566 pp.

Notes: Abstract. Two new viruses, goldfish virus-1 (GFV-1) and goldfish virus-2 (GFV-2), were isolated from swimbladder cultures prepared from healthy Carassius auratus (L.). The isolates were sensitive to pH, heat, and chloroform but stable to freezing and thawing. Treatment with IUDR, Feulgen staining and acridine orange staining indicated that the viruses contain double-stranded DNA. Electron microscopy revealed enveloped virions of 180 nm enclosing icosahedral capsids of 120 nm. Viral growth was studied by light and electron microscopy and infectivity assays. Virions were assembled in cytoplasmic inclusion bodies and apparently released through cell degeneration. Infectious virus was first produced at 24 h post-infection. Progressive cytopathology was observed leading to cell ghosts by 96 h post-infection. The results suggest tentative placement of the viruses in the family Iridoviridae.

Notes: Abstract. The in vitro bacteriostatic activity of a combination of sulphadiazine (SDZ) and trimethoprim (TMP) against representative bacterial fish pathogens was studied. In general, the minimum inhibitory concentrations (MIC) of TMP were much smaller than comparable values for SDZ. When SDZ and TMP were combined, the bacteriostatic effect was enhanced in a synergistic manner for all bacteria tested except Pseudomonas fluorescens. The synergism was most remarkable in the case of bacterial strains that were sensitive to the action of SDZ alone. This degree of synergism was observed when the ratios of SDZ and TMP in combination were in the range of 1:1 io 5:1. The MIC of SDZ plus TMP combined was less than 3.13 μg/ml for SDZ resistant strains and as low as 0.2 to 0.78 μ/ml for SDZ sensitive strains. The possible usefulness of a combination of these two compounds for therapy or control of bacterial fish disease is discussed.

Notes: Abstract. Trophonts of Pisdnoodinium pillulare (Schäperclaus, 1954), a common ectoparasite of freshwater aquarium fish, are attached to host cells by means of a specialized structure, the attachment disc. Unlike other dinoflagellate genera parasitic on fish and invertebrates, this disc features nail-like organelles, the rhizocysts. Head-parts of the rhizocysts are inverted in separate compartments, rhizothecas, in the sole of the disc while their long shafts are firmly embedded in the cytoplasm of cells of the host epidermis or gill epithelium. The attachment inflicts a serious injury on the host cells which may ultimately be destroyed. Rhizocysts originate in the subnuclear cytoplasm from where they migrate into the attachment disc. There are other specialized organelles and inclusions; fibrous vesicles, membraneous bodies, striated tubular bodies and paracrystalline bodies. Pisdnoodinium has well-developed chloroplasts. While its cytological adaptations indicate a nutritional dependence on the host, there is no evidence of ingestion of host-derived particulate material. Pisdnoodinium may derive an essential part of its nutrition from photosynthesis.