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The various pathology of female mice reproductive system with the mutation smoky (smk) was observed. This recessive autosomal gene dilutes the black colour of hair to the light smoky. The cases with ovarian agenesis and dysgenesis of the mutants WR-smk/smk and B6-smk/smk without the germ cells or with the acute deficit was discovered. The oogenesis pathology of the mutants B6-smk/smk was revealed by the block of oocyte growth and their nucleus development in the growing nonatretic follicles leading to the female infertility. Endogene causes of the female hypogonadism and infertility associated with the mutation smoky are proposed.

Recombinant H-2 haplotype of mouse strain B10.D1(R108)/Y (symbol R108) obtained in experiments with skin grafting in the course of developing the CR B10.D1/Y strain (strain DBA/LacY--the donor of H-2q) was studied. Strains with recombinant H-2 haplotypes a, h2, g1, i3, i5, i7, m, y1 were used. Alleles of different H-2 (K, I, D) regions were determined according to the presence or absence of genetic complementation in the F1 test with skin grafts. R108 recombinant was studied by serological methods with panel of anti-H-2 sera. Anti-H-2Kb (H-2.33) and anti-H-2Dq (H-2.30) monospecific antisera were used in microcytotoxicity test and in absorption experiments in vitro. It was concluded that crossing over between H-2b and H-2q chromosomes, which led to formation of recombinant H-2 haplotype of R108 mice, occurred at I region, between IA and IC subregions. The H-2 complex of R108 line has KbIAbIJ?IE?ICqSqDq alleles. bq1 symbol was proposed for the H-2 haplotype of B10.D1(R108)/Y strain.

Genetic control over sensitivity to experimental adjuvant arthritis was studied in different strains of inbred, recombinant and congeneic mice. The development of adjuvant arthritis is genetically regulated, with sensitivity to adjuvant arthritis controlled by H62-complex. The genes controlling sensitivity to adjuvant arthritis were shown to be located in (S-region) H-2 system.

Specific suppressor T cells (SSTC), primed in vivo with H-2 antigens, have been shown previously to inhibit DNA synthesis in the one-way, three-cell mixed lymphocyte reaction (MLR) provided that (a) the stimulator cells bear the priming H-2 antigens, and (b) the responder cells possess IC + S regions homologous to those of the SSTC. Anti-B10.A B10.A(2R) SSTC (anti-Dd) and anti-A.AL A.TL SSTC (anti-Kk) are shown here to be able to inhibit the DNA synthesis triggered in MLR, not only by the corresponding antigens, Dd and Kk, respectively, but also by irrelevant, third-party H-2 and Mls products provided that the corresponding and third-party antigens are presented on the same stimulator cell. If stimulator H-2 regions, whose products interact with SSTC and responders, are located on different stimulator cells within the particular MLR, SSTC activity is not elicited. Participation of cytotoxic T lymphocytes in DNA-synthesis suppression is ruled out. Direct contact or location of the inhibited responder cell very close to SSTC is considered to be required for the development of SSTC activity.

The optimal conditions are found for in vivo irradiated lymphocyte induction of high cytotoxic T lymphocytes (CTL) specific to the H-2Kb molecule with a subsequent monoculture differentiation. B10.D2(R101) CTL have a pronounced excess as compared to B10.A (4R) CTL, with respect to lysis intensity of the same target cells (TC), and requires a lower term of the monoculture incubation in spite of their specificity to the same H-2Kb molecule. As the susceptibility of TC for CTL lysis is higher (M phi as compared to EL-4 thymoma cells), CTL are much more inactivated with the monoclonal antibodies to Lyt-2 and Lyt-3 antigens without complement. Anti-H-2Kb CTL differentiated in the monoculture cross react to TC bearing third-party H-2 molecules (Kk, Dq, Dk). Unlike a stable CTL adherence to the donor M phi monolayer, nonspecific CTL adherence to the syngeneic M phi monolayer declines in the presence of EGTA, and as a result of repeated detachment of lymphocytes. The findings give rise to study receptor affinity expressed on the in vivo induced CTL surface, the CTL receptor monoclonal antibody and the CTL differentiation factor.

Spotted sterile male - a new mutation in mice is described (tentative symbol Ssm). White spotting on the belly, legs and tail as well as sterility in heterozygous males Ssm/+ of the B10.M strain are caused by autosomal semidominant gene Ssm. The gene is localized on the 5 chromosome: the frequency of recombination between Ssm and go is 13.6 +/- 1.6%; Ssm is closely linked to Wv. The diheterozygotes Ssm+/+Wv are darkeyed white sterile mice. The deficiency of spermatogenic epithelium cells, emptyness of seminiferous tubules as well as interstitial tissue overgrowing occurred in the testis in sterile males Ssm/+ of B10.M. The fertile hybrid males Ssm/+ are obtained in outcrossing of females Ssm/+ of B10.M with males of YT/Y, CBA/CaY, DBA/2JY, A.CA/Y strains.

Spleen T-cells of mice immunized intravenously with gamma-irradiated allogenic lymphoid cells at a high dose are shown to inhibit the activation of DNA synthesis in MLC. The difference in the whole H-2 complex or in either of its K, I, D regions or I-subregions is required and sufficient for induction of specific T-suppressors. The magnitude of the suppression is a sum of suppressive reactions induced separately by each of the immunizing complex components, the products of one of the H-2 regions (subregions) as shown by changing the source of stimulator cells in MLC. These components have a similar impact in the suppressor activity. Unlike the other T-cell subclasses, T-suppressors react only to serologically defined mutant antigens of the Dd allele, but do not react to the mutants of the Kb allele and do not discriminate Kb and Kba antigens. Specific suppressor T-cells recognize the I-subregion products without their linkage to the K/D region products. For elicitation of the suppression two conditions are required and sufficient: a) direct contact of immune suppressors with the corresponding antigenic determinant; b) the subregion IC identity between the suppressors and the responder cells. Intensity of suppression is reduced if F1 hybrid responder cells interact with parent suppressor cells, but is not reduced in the inverse arrangement of the experiment. Specific suppressor T-cells are presumed to differ from other subclasses of T-cells and to be similar to B-cells relatively to the nature and multiplicity of recognizable individual products of the H-complex and to narrow specialization of reactive clones.

With the help of antibodies-containing serum reacting with the thymus reticulum epithelial cells components by immunofluorescence method the thymus parenchimal tissue in mice with functional and pathological involution has been detected electively. In spite of large thymus changes in hairless mice with the mutation of gene hrrhy in 14-th chromosome of B10.R109/Y animals the basal cells antigen in epithelial reticulum has been preserved. It permits to estimate thymus involution level of this organ. Low lymphocyte content in the thymus of C57BL/6 mice is accompanied by total decamouflage of the epithelium. The functional thymus involution of pregnant mice is characterized by the luminescence of large number of epithelial cells, the restoration of organ after the delivery--by their few number. The elective detection of thymus epithelium many serve as additional test for the estimation of functional and pathological involution level.

The differences in the H-2 system between mice of the C57BL/10 (H-2b) and 129 (H-2bc) were studied in crosses B10.D (H-2d) or recombinant strains R106 (KbSbDda), R107 (KbSbDd) and R101 (KdSdDb) with strains 129 or B10.129 (6M). Most skin grafts from the strain C57BL/10 were rapidly rejected in the F1 hybrids derived from the crosses with strains B10.D2, R106 and R107, thus indicating to the effect of the relatively strong H-locus either in the D region or in the Tla region of H-2b. Chronic responses were observed in the F1 hybrids derived from crosses with the strains R101 and R103, indicating to the weak effect of another locus. However all the skin grafts survived in the F1 hybrids with the strain HTG (KdSdDb). This weak H gene is apparently located to the left of the crossover in the recombinants R101 and R103, but to the right of the crossover in the HTG, i. e. between the loci Slp and H-2D. The new locus is tentatively designated as H-2W.

Specific d anti-b and b anti-a suppressor T cells induced by intravenous injection of mice with gamma-irradiated allogenic lymphoid cells, are not a homogenous population of cells as shown by their selective absorption on macrophage monolayers of various H-2 haplotypes. This is proved by separation of suppressor T cells to two subpopulations, at least, each of them being able to react with the products of only one (K or D) end of the H-2 complex. Moreover, the fine specificity study of d anti-b suppressor T cells enriched by elution from macrophage monolayers of different H-2 haplotypes, demonstrated these suppressors to represent a set of narrow-specific clones, each of them carried receptors reactive in a selective fashion with a particular determinant of the H-2 molecule irrespective of this linkage with other products of the H-2 complex. Two such clones reactive with H-2a and H-2f third-party antigens, respectively, were isolated by elution from the corresponding cell monolayers, each of them accounted for about 1.5% of the total d anti-b suppressor population. These data are discussed in the light of differences of suppressor T cells from other T cell subclasses and their resemblance to B cells with respect to the clonal structure and the receptor specificity.

The injection of 6 x 10(9) sheep red blood cells (SRBC) to mice suppressed the delayed type hypersensitivity (DTH) in situ and activated spleen T cells which prevent sensitization of syngeneic recipients. Similar effect was obtained when suppressor cells induced in F1 hybrids were transferred to parental mice. Suppression was also reached in allogeneic strain combination if suppressor cells of donors and recipients shared the major histocompatibility complex (MHC). Studied performed with recombinant and mutant strains revealed that the prerequisite for interaction of DTH suppressors and effectors was the identity of K-region of MHC. Passive transfer of DTH to SRBC was also possible if donors and recipients were identical in K-region of MHC. It is believed that interaction between DTH suppressors and effectors is restricted by a region of MHC whose product takes part in antigen representation.

The MC-resistant specific suppressor T cells that inhibit DNA synthesis and CTL generation in MLC were induced in vivo by gamma-irradiated allogeneic lymphoid cells in a large dose. MLRs were inhibited only slightly when triggered by third-party cells, even neighbouring with the corresponding stimulators. Unlike the irradiated cells, intact allogeneic lymphoid cells induced a mixture of macrophage-like and T-cell suppressors with a pronounced non-specific component of the action. Syngeneic cells induced low active non-specific suppressors of macrophage type only. The suppression was not due to a cytotoxic effect, since specific T suppressors differed from CTL by conditions of induction and high sensitivity to gamma-irradiation and from CTL precursors by high sensitivity to CY and HC. The specific T-suppressors could be selectively removed by adherence to a macrophage monolayer of the corresponding donor. The subsequent elution of adherent lymphocytes with pronase resulted in enrichment of specific T suppressors by a factor of 30 and 2.6, as judged by reduction in the number of lymphocytes required for 50% inhibition of DNA synthesis and 33% inhibition of CTL generation, respectively. The high specificity of this enrichment is shown by using both syngeneic monolayers for fractionation and third-party stimulators in MLR for testing and by disappearance of slight non-specific suppression caused by non-fractionated suppression. Complete inactivation of the eluted suppressors with anti-Thy-1.2 and anti-T antibodies, their resistance to anti-Mls antibodies, carrageenin, and carbonyl iron, together with the data of autoradiographic study and total DNA synthesis in the population indicate that the eluted highly specific suppressors represent mainly DNA-synthesizing large and medium T lymphocytes.

Differences in the generation of anti-H-2Kb wild-type specific cytotoxic T lymphocytes (CTL) and specific suppressor T cells (SSTC) were investigated in H-2Kbm mutant mouse strains. To this end, optimal conditions for in vivo induction of highly active CTL in this mutant system were found and the T-cell origin of CTL and SSTC was confirmed using the anti-Thy-1.2 monoclonal antibody G4. Unlike the CTL, which were generated in vivo by any of the mutant strains tested (bm1, bm3, and bm4), the SSTC were only produced by bm3, whose H-2Kbm3 antigen, in contrast to the other H-2Kbm molecules, differs from wild type by serologically defined determinants. The high activity of anti-wild type bm4 CTL induced in vivo contrasted with a low activity of such CTL induced in mixed lymphocyte culture (MLC). This appeared to be the property of bm4 only, but not of bm1 or bm3, and it was reproduced in the reciprocal system B10 anti-bm4. CTL generation could be restored in the MLC by the addition of concanavalin A supernate or a mixture of bm4 and bm12 stimulator cells. Three of the six in vivo induced and in vitro propagated bm3 anti-B6 CTL clones demonstrated selective cross-reactivity to only one of the third-party H-2K molecules used, either Kk, Kd, or Kbm4. The present results indicate that (a) the SSTC and antibody recognize similar H-2Kb epitopes; (b) the H-2Kb epitopes recognized by the CTL and SSTC are not identical; (c) the genetic control of CTL generation in vivo is distinct from that in MLC, and (d) the affinities of antigen-specific receptors on T-cell clones of the same specificity may be different, leading to their individual cross-reactivity patterns.

C57BL/6 (H-2b) mice were immunized once intravenously by irradiated (1500 rad) spleen cells of mutant strains B6.C-H (z1)Y (H- 2bm1 ) and BB. M505Y (H- 2bm3 ), by the cells of allogeneic strain B10.D2 (H-2d) (in positive control) and syngeneic strain C57BL/6 (in negative control). It was demonstrated that MIF-producing T cells can react to mutant H-2 antigens and that this reaction is revealed on the 1st day after intravenous immunization in vivo. It is suggested that MIF-producing T cells can participate in the immunological surveillance of slight changes in the major histocompatibility complex antigens through mobilization of macrophage effector functions.

Two antisera to Ia antigens, products of the H-2 complex I-Cd and I-JkEk subregions, respectively, have been obtained by immunization of the F1 hybrids of recombinant strains of mice. These antisera are shown to display a 50% cytotoxic effect in vitro, in the presence of complement, upon lymphocyte populations immune to the H-2-complex antigens and enriched for specific suppressor T cells (SSTC) by fractionation on a monolayer of target cells. The specificity of anti-Ia cytotoxins is shown by cross-antibody absorption with T and B cells of mice originating from the recombinant H-2 haplotypes and bearing either particular I-Cd, I-Jk and I-Ek antigens, or their combinations. Anti-I-Cd cytotoxins were found to react with both B and T cells, but at a different rate, and the anti-I-JkEk serum contains two antibody types directed to I-Ek and I-Jk products, respectively, the latter being able to react preferently with T cells. Although both antisera do inactivate the in vitro SSTC function in the presence of complement to a similar degree, the inactivating action of the anti-I-Cd serum, but not that of the anti-I-JkEk serum, occurs without complement. SSTC are shown to bear both Ia-antigens, I-J and I-C, as shown by both inactivation of the anti-suppressor effect of the antisera absorbed with spleen cells of different H-2 origin, and variation of the H-2 origin of SSTC pretreated with the intact antisera. It is suggested that these two markers, located on the same SSTC, function differently in SSTC immune to the H-2 antigens, and I-C antigen expression on the SSTC surface is presumed to be required for their interaction with the inhibited responder T cells proliferating in MLC.

The long duration of the estrous cycle, the absence of the ovulated oocytes was observed in histomorphological studies of estrous cycles and ovaries in hr rhY-/hr rhY-mutants of B10-hr rhY mice strain. The absence of mutation division, ovulation and corpora lutea in ovaries was found in contrast to normal isogenic B10 females. The extragonadal cause of hairless mutant female infertility was shown. The effective method of mutant strain B10-hr rhY reproduction by using the transplantation of homozygous hairless female ovaries to isogenic recipients (+/+ or +/hr rhY) was proposed.

In vivo induced d anti-b spleen cytotoxic T-lymphocytes (CTL) display the cross lysis of H-2f and T-2a target cells (TC) which is about 4 to 6 per cent of the H-2b TC lysis as judged by comparison of CTL doses required for the same lysis. d anti-b CTL fractions cross-reacting (CR) to H-2f and H-2a TC are not identical and can be separated one from another by a selective adherence to the corresponding TC monolayer. The preliminary CTL absorption onto the syngeneic TC monolayer and the 3-step elution of the adherent CTL by pronase in two concentrations and EDTA gave rise to the optimizations of the CTL enrichment conditions and to the fractionation of CTL on the basis of stability of their contact with monolayer cells. The eluted CR d anti-b CTL fractions were found to destroy H-2b TC much stronger than TC of the CR strains from which CTL were eluted, to cross-react to the irrelevant CR antigen and to destroy H-2b TC as much as the CTL eluted from the H-2b monolayer. Besides, equalization and even invertion of the activity of CTL fractions was observed with respect to H-2b TC if CTL were eluted from CR strain monolayers. CTL receptors are suggested to be directed to the single CTL-determinant of an H-2 antigen and to be unable to seen particular serologically defined common specificities of the same antigen. The CTL cross reaction is supposed to be the variability in the unified CTL receptor complementarity (affinity); the less complementary (rigid) receptors being able to accommodate one of particular CR H-2 antigens.

The ability of T-cells producing the migration inhibition factor (MIF) to recognize point mutations of the K-end of the H-2 complex was revealed in a primary mixed lymphocyte culture. Nonimmune C57BL/6Y (H-2bm) lymphocytes produce MIF in response to the H-2 antigens of B6..C-G (zI)/Y (H-2bm1) and B6.M505/Y (H-2bm3) mutant strains. Similarly lymphocytes of the mutant strains react to the H-2b wild-type and reciprocally to each other. T-cell dependence of these reactions was shown by their abolition after pretreatment of responder cells with Thy-1.2 antiserum in the presence of complement. The ability of MIF-producing cells to recognize serologically unrecognizable H-2bm1 and H-2bm3 mutant antigens demonstrates high immunologic specificity and sensitivity of the MIF-reaction.

The immune response of (CBA X M523)F1 mouse lymphocytes to the antigen (sheep red blood cells) in CBA mice after lethal irradiation was studied. When irradiation, cell transfer and antigen test-injection were performed at the same day the graft activity was inhibited, in comparison with syngeneic system. The activity of donor cells restored on increasing the interval between these processes up to 3 days. Retransplantation of the spleen cells from recipients to irradiated CBA and F1 mice revealed viability of transplanted cells and lack of their readaptation to insyngeneic microenvironment. The recipient's resistance could be specifically overcome by the preinjection of mouse cells combined with cyclophosphamide or without it. The results obtained allow a conclusion that genetic parental resistance of CBA mice to F1 mouse cells is due to the recipient immunocompetent cells which are inactivated 3 days following irradiation. They do not produce cytotoxic effects with respect to donor cells, blocking, however, their activity for some time.

Lymphocytes of mice F1 (CBA X M523) and F1 (A X M523) transplanted to 1000 R irradiated CBA or A mice responded to the test antigens--SRBC or S. typhi Vi-antigen--by formation of 100--1000 times less antibody forming cells than in syngeneic recipients. An intermediate result is achieved when the lymphoid cells are transplanted to the irradiated M523 mice. Lymphocytes of mice F1 (A X CBA), F1 (CBA X C57Bl/6), or F1 (A X A.CA) developed a similar immune response in the irradiated syngeneic mice and in both parental lines. The ability of parental line M523 to respond to SRBC was the same as in the other lines studied when examined in situ or in adoptive transfer experiments. The stem hemopoietic cells of mice F1 (CBA X M523) develop in the spleen of CBA mice 2--2.5 times less hemopoietic colonies than in the spleen of syngeneic animals. A conclusion was drawn that mutation M523 in CBA mice inhibited the proliferation and differentiation of hemopoietic and lymphoid cells in the irradiated nonsyngeneic recipients.

Hz1 and M505 are two mutants affected the Kend of H-2 major histocompatibility system in mouse. Inability of M505 to complement Hz1 is considered as an evidence that the same gene is altered by both mutations. The gained antigens of two mutants can cross-react in vivo, and the lost antigenic determinants are not identical as revealed in skin grafting tests with presensitized recipients. The antiserum ASY-0,15, (d X a) anti-i, could differentiate between normal and mutant cells as well as between both mutants in absorption tests. These results show that some haptenic determinants are changed in the Hz1 and M505 mutations together with changes of histocompatibility determinants.

Lymphocytes B10D2 anti-B10 (anti-KbIbDb) produce MIF when incubated with spleen cells of the recombinant KbIb-bearing R107 and Db-bearing R101 lines, possessing either public or private H-2 specificities of the immunizing complex. Cross reactivity revealed in the reaction with "unrelated" lines one week after immunization by allogeneic tumour disappeared during the second week; simultaneously the intensity of the reaction to KbIb and Db region products changes. The cross reaction of anti-Kb MIF producers obtained one week after immunization with cells of Kba and Kbd mutants disappeared two weeks after immunization: the migration inhibition which occurred one week after immunization turns to stimulation during the second week, while the degree of inhibition in the reaction with B10 cells remains unchanged.

The nature of cells responsible for the genetic resistance of lethally irradiated CBA mice to lymphocytes of (CBA x M523)F1 hybrids was studied. Preirradiation of the hosts was shown to abolish the resistance. The latter was generally recovered by syngeneic thymocytes or splenocytes while embryonic liver and bone marrow cells or splenocytes treated with anti-Thy-I serum plus complement before injection into host were ineffective. It is postulated that some cells with T cell characteristics are responsible for the phenomenon of parental resistance. These cells differ in several respects from T cells that mediate the transplantation immunity and from M cells that control other forms of the genetic resistance.

A new spontaneous mutation of the H-2b haplotype was found in skin graft tests with BC3 mice derived from B10.R111 (71NS) and C57BL/10SnY outcrossing. The mutation site localized in the F1 test in the H-2Kb gene is nonidentical to and noncomplementary with bm1, bm3, bm4 mutations. The novel mutation is maintained as B10.R111-H-2bm25 strain.

The differences in the generation of specific suppressor T cells (SSTC) against H-2Kb wild type were investigated in H-2Kbm1, H-2Kbm3 and H-2Kbm4 mutants. Anti-Kb SSTC were produced only by bm3 mutant and F1(BALB/c X bm3) hybrid. T-cell nature of SSTC of bm3 mutant was confirmed by anti-Thy 1.2 monoclonal antibodies described in the same study.

(CBA X M523)F1, (A X M523)F1 and M523 lymphocytes grafted into lethally irradiated CBA or A mice temporarily lose their capacity to respond to test antigens (SRBC, Vi-antigen, S. typhi). Immunoresponsiveness of F1 cells is affected to a lesser degree in lethally irradiated M523 mice. Depression of response is absent in the CBA leads to F1 combination, in the syngeneic combination and in CBA mice whch have received transplanted cells from F1 hybrids which do not share the M523 mutation. The number of hemopoietic (CBA X M523)F1 colonies was also reduced in CBA mice. Resistance of CBA mice to lymphoid (CBA X M523)F1 cells develops 18 days after birth. It can be reduced by additional recipient preirradiation or preinoculation with (CBA X M523)F1 spleen cells. The abrogated resistance can be partially restored by CBA spleen cells. The activity of (CBA X M523)F1 lymphocytes passaged through CBA spleen is restored in syngeneic F1 secondary recipients but inhibited again in the CBA secondary recipients. These results are consistent with the suggestion that resistance of lethally irradiated CBA mice to hemopoietic and lymphoid (CBA X M523)F1 cells is mediated by immunologically competent, radioresistant recipient cells rapidly reacting to transplantation antigens coded by the mutant H-2Kka allele. These cells temporarily suppress the functional activity of transplanted cells but do not eliminate them.

Two antisera to Ia antigens, products of the H-2 complex I-Cd and I-JkEk subregions, respectively, have been obtained by immunisation of the F1 hybrids of recombinant strains of mice. These antisera are shown to display the 50 per cent cytotoxic effect in vitro in the presence of complement upon lymphocyte populations immune to the H-2 complex antigens and enriched for specific suppressor T cells (SSC) by fractionation on the monolayer of target cells. The specificity of anti-Ia cytotoxins is shown by the cross antibody absorption with T- and B-cells of mice originated from the recombinant H-2 haplotypes and bearing either particular I-Cd, I-Jk and I-Ek antigens, or their combinations. Anti-I-Cd cytotoxins are found to react with both B and T cells at a different rate, and the anti-I-JkEk serum contains two antibody types directed to I-Ek and I-Jk products, respectively, the latter being able to react preferently with T cells. Although both antisera do inactivate the in vitro SSC function in the presence of complement at a similar degree, the inactivating action of the anti-I-Cd serum, but not that of the anti-I-JkEk serum, occurs without complement. SSC are established to bear both Ia-antigens, I-J and I-C on the same cell, as demonstrated by the cross antibody absorption and variation of the H-2 origin of SSC. These two markers are suggested to function differently in the SSC immune to the H-2 antigens and the I-C antigen expression on the SSC surface is presumed to be required for their interaction with the inhibited responder T cells proliferating in MLC.