Major Transplantation Antigens Research Articles

Differential expression of MHC class I antigens on the placenta of the rat. A mechanism for the survival of the fetal allograft.

In some mating combinations in rats, there is a maternal antibody response to the maternal antigenic components of the placenta without any previous immunization of the mother. The highest response occurs in the WF (u) female mated to the DA (a) male, and it is against a unique MHC-encoded class I antigen, the Pa antigen, and not against the major allele-specific transplantation antigen of the DA strain, RT1.Aa. The development of mAbs to the Pa and Aa antigens allowed us to localize these antigens on the placenta and to explore the reason for the differential antibody response to them using immunohistochemical and biochemical techniques. Both antibodies reacted with the WF X DA placenta and stained the endovascular and interstitial trophoblast of the decidua, the basal trophoblast, Reichert's membrane, and the yolk sac epithelium, but they did not stain the labyrinthine trophoblast. Blocking studies showed that each antibody reacted with a separate molecule in the placenta. Anti-class II mAbs reactive with the a or u haplotype did not stain the WF X DA, DA X DA, or WF X WF placenta; hence, there are no class II antigens in the placenta. Electron microscopic studies of the semiallogeneic WF X DA placenta using the immunogold technique with both single- and double-labeling showed that only the Pa antigen was expressed on the surface of the basal trophoblast, but that both the Pa and Aa antigens were in the cytoplasm of these cells; neither antigen was found in the labyrinthine trophoblast. By contrast, the placenta from the syngeneic DA X DA mating expressed both the Pa and Aa antigens on the surface of the basal trophoblast as well as in the cytoplasm; neither antigen was found in the labyrinthine trophoblast. These observations were quantified morphometrically using electron photomicrographs of single-labeled tissues. Both the Pa and Aa antigens isolated from the plasma membrane of lymphocytes have heavy chains of 46 kD, but those antigens isolated from the plasma membrane of basal trophoblast cells have heavy chains of 43 kD. Based on densitometric measurements of autoradiographs, the Pa/Aa ratio in the basal trophoblast membrane is 23.5, whereas it is 0.46 in lymphocyte membranes. These studies show that there is differential regulation of the expression of class I antigens on basal trophoblast cells in semiallogeneic pregnancies, but not in syngeneic pregnancies, such that the major allele-specific transplantation antigen is scarcely expressed on the surface of the basal trophoblast.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of low or high doses of short wavelength ultraviolet light (UVB) on Langerhans cells and skin allograft survival.

Since Langerhans cells (LC) are normally the only cells within the epidermis to express the class II major histocompatibility complex (MHC) transplantation antigens, depletion of LC could be expected to prolong skin allograft survival by reducing the antigenic disparity between host and recipient. To assess this hypothesis, donor C57BL mouse shaved dorsal trunk or tail skin was exposed to high (200 mJ/cm2) or low (40 mJ/cm2) doses of short wavelength ultraviolet light (UVB) before grafting on to the thorax of BALB/c mouse recipients of the same sex. These strains have different major and minor transplantation antigens. The effects of UVB treatments on LC were determined by electronmicroscopy. Skin grafted 1-14 days following a single high dose of UVB irradiation was ultrastructurally depleted of LC and survived significantly longer than unirradiated skin before being rejected. After a 21-day interval between exposure and grafting when LC were again present in the epidermis there was no significant difference between treated and control graft survival. Exposure to low dose UVB irradiation only significantly increased graft survival for skin transplanted 1-3 days after irradiation; skin grafted 4 days following irradiation survived for a similar period to unirradiated control skin grafts. Electronmicroscopy showed that the low UVB dose did not deplete LC from the epidermis. We conclude that after low dose UVB treatment the class II MHC antigens on the LC plasma membrane were lost temporarily, thus prolonging graft survival, but when the plasma membrane antigens were re-expressed graft survival returned to normal.(ABSTRACT TRUNCATED AT 250 WORDS)

A trans-acting mechanism represses the expression of the major transplantation antigens in mouse hybrid thymoma cell lines.

We have fused an H-2- thymoma (BM5R.9) with an H-2+ thymoma (BW5147) and have found that many of the resulting hybrids exhibit an H-2- phenotype. In several hybrids that were analyzed in detail, this phenotype is related to the absence of steady-state H-2 mRNA and shows some instability, possibly related to the loss of chromosomes in segregants. We conclude from our studies that BM5R.9 cells display a trans-acting mechanism that can repress the expression of H-2 antigens, and that the gene(s) causing the repression are not located on chromosome 17. This mechanism is not sufficient to explain the H-2- phenotype of BM5R.9, for which an additional, cis-acting process, must be postulated. We discuss these results in the context of the regulation of expression of the major class I transplantation antigens.

Biological role of major transplantation antigens in T cell self-recognition.

The proposal is made, illustrated and supported by experimental evidence that T cell-mediated immunopathology triggered initially by low- or non-cytopathic infectious agents may cause diseases, susceptibility to which is linked to the major histocompatibility gene complex.

Production of C3W mouse strain congenic to C3H/HeN at the newly defined minor H-42 histocompatibility locus.

Microbiology and ImmunologyVolume 30, Issue 9 p. 939-943 Original ArticleFree Access Production of C3W Mouse Strain Congenic to C3H/HeN at the Newly Defined Minor H-42 Histocompatibility Locus Hidehito Kato, Hidehito Kato Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this authorHiromichi Ishikawa, Hiromichi Ishikawa Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this authorTomoo Hino, Tomoo Hino Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this authorHiroko Suzuki, Hiroko Suzuki Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this authorKazuhisa Saito, Kazuhisa Saito Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this author Hidehito Kato, Hidehito Kato Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this authorHiromichi Ishikawa, Hiromichi Ishikawa Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this authorTomoo Hino, Tomoo Hino Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this authorHiroko Suzuki, Hiroko Suzuki Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this authorKazuhisa Saito, Kazuhisa Saito Department of Microbiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, 160Search for more papers by this author First published: September 1986 https://doi.org/10.1111/j.1348-0421.1986.tb03024.xCitations: 1AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat REFERENCES 1Bechtol, K.B., and McDevitt, H.O. 1976. Antibody response of C3H ↔ (CKB x CWB) F1 tetra-parental mice to poly-L-(Tyr, Glu)-poly-D, L-Ala-poly-L-Lys immunization. J. Exp. Med. 144: 123– 144. 2Ishikawa, H., Kubota, E., Suzuki, H., and Saito, K. 1985. The target minor H antigen for F1 cytotoxic T lymphocytes induced by Igh-congenic parental spleen cells is coded for by gene linked to H-2. J. Immunol. 134: 2953– 2959. 3Ishikawa, H., Suzuki, H., Hino, T., Kubota, E., and Saito, K. 1985. In vivo priming of mouse CTL precursors directed to product of a newly defined minor H-42 locus is under a novel control of class II MHC gene. J. Immunol. 135: 3681– 3685. 4Juretic, A., Vucak, I., Malenica, B., Nagy, Z.A., and Klein, J. 1984. H-41, a new minor histocompatibility locus. I. Histogenetic analysis. J. Immunol. 133: 2950– 2954. 5Klein, J., Figueroa, F., and Klein, D. 1982. H-2 haplotypes, genes, and antigens: second listing 1. Non-H-2 loci on chromosome 17. Immunogenetics 16: 285– 317. 6Klein, J., and Herzenberg, L.A. 1967. Congenic mouse strains with different immunoglobulin allotypes. I. Breeding schema, histocompatibility tests, and kinetics of γG2a-globulin production by transferred cells for C3H. SW and its congenic partner CWB/5. Transplantation 5: 1484– 1495. 7Simpson, E., and Gordon, R.D. 1977. Responsiveness to HY antigen. Ir gene complementation and target cell specificity. Immunol. Rev. 35: 59– 75. 8Zinkernagel, R.M., Althage, S., Cooper, S., Kreeb, G., Klein, P.A., Sefton, L., Flaherty, L., Stimpfling, J., Schreffler, D.C., and Klein, J. 1978. Ir-genes in H-2 regulate generation of anti-viral cytotoxic T cells. Mapping to K or D and dominance of unresponsiveness. J. Exp. Med. 148: 592– 606. 9Zinkernagel, R.M., and Doherty, R.C. 1979. MHC-restricted cytotoxic cells: studies on the biological role of polymorphic major transplantation antigens determining T-cell restriction-specificity, function, and responsiveness. Adv. Immunol. 27: 51– 177. Citing Literature Volume30, Issue9September 1986Pages 939-943 ReferencesRelatedInformation

Asynchronous regulation of mouse H-2D and beta-2 microglobulin RNA transcripts.

The major transplantation (or H-2) antigens in the mouse are cell-surface glycoproteins composed of a heavy chain and a light chain, the beta-2 microglobulin (beta 2m). The expression of these proteins is regulated during development. Embryonic cells at early stages of development do not express these proteins. On the other hand, these molecules are present on the surface of all adult somatic cells. We investigated whether the expression of both chains was coordinately regulated. Using specific single-stranded DNA probes in an S1 nuclease analysis, we compared the relative amounts of H-2D and beta 2m transcripts in normal tissues, in transformed cells, and during embryonic development. Our results show that (1) the steady state level of beta 2m transcripts varies from one adult organ to another, while that of H-2D transcripts stays approximately the same; (2) upon transformation, the amount of H-2D-specific mRNA increases drastically, while the beta 2m mRNA level remains constant; (3) whereas the quantity of beta 2m mRNA increases during early development, the amount of H-2D mRNA remains at a very low level. These data suggest that the regulation of H-2D and beta 2m genes are not identical and that their activation during development is not synchronous.

Expression of class I genes in the major histocompatibility complex: identification of eight distinct mRNAs in DBA/2 mouse liver

The mouse H-2 multigene family includes the genes coding for the major transplantation antigens and for genes located in the Qa-TIa region. We have studied a collection of class I cDNA clones made from liver mRNA of DBA/2 mice (H-2d haplotype) and found that at least six distinct class I genes are transcribed, including three genes of the Qa-TIa region. Two of these six genes each yield two distinct mRNAs, resulting from alternate splicing. Altogether, liver cells may express at least eight distinct class I polypeptides, of which three might be secreted, while one may be a new presumptive nonpolymorphic surface antigen.

Alternative splicing in the 5' moiety of the H-2Kd gene transcript.

The H-2Kd gene, which encodes a mouse major transplantation antigen, was transfected into L TK- mouse fibroblasts. Two transcripts of the gene were detected by S1 nuclease mapping analysis. They correspond to two previously characterized cDNA clones isolated from DBA/2 mouse liver RNA, leading to the conclusion that the H-2Kd gene gives rise to two distinct transcripts through an alternate use of splicing sites. The non-canonical RNA potentially encodes a so far undescribed H-2Kd-like molecule. It is present in all tissues tested (liver, spleen, thymus, kidney) albeit in lower amounts (approximately 10-fold less) than the canonical RNA coding for H-2Kd.

Suppressor T cell recognition of major and minor histocompatibility alloantigens: selected suppression of MHC-directed responses by minor alloantigen Ts.

The induction of suppression by i.v. administered alloantigens in the murine host was analyzed as a model of the possible effects of blood transfusion on transplant survival. The results indicated that suppressor T cells (Ts) specific for minor histocompatibility alloantigens could be readily induced by the i.v. presentation of minor alloantigen-disparate spleen cells. In contrast, similar priming with cells differing solely at the H-2 major histocompatibility complex stimulated only positive T cell immunity, with no evidence of suppression. The induction of H-2 directed Ts activity could be accomplished only by i.v. priming with major plus minor incompatible donor cells, suggesting that suppressor cell recognition of minor alloantigens may have facilitated the generation of Ts against H-2-encoded major transplantation antigens. A role for minor histocompatibility antigens in the regulation of H-2-specific immunity at the effector level was also indicated. Ts induced by i.v. pretreatment with minor antigen-disparate donor cells not only suppressed the delayed-type hypersensitivity (DTH) response to the relevant minor alloantigens, but also inhibited DTH against unrelated H-2 alloantigens introduced during subsequent intradermal immunization. Suppression of H-2-directed T cell reactivity was specific in that the presence of the Ts-inducing minor alloantigens was also required and occurred only when the minor and unrelated major alloantigens were presented within the same inoculum, if not on the same cell surface. The capacity of Lyt-2+Ts or Ts-derived suppressive factors specific for one set of cell surface molecules to modulate responses to an unrelated group of surface antigens does not appear to represent a general phenomenon, because similar suppression of immunity to unrelated tumor-specific transplantation antigens by minor-specific Ts was not observed. These results are discussed with respect to the possible mechanism of H-2-directed suppression and the role of the I region in Ts recognition of antigen.

Failure of cyclosporin-A to induce immunological unresponsiveness to nerve allografts

Although some allografts bearing major and minor transplantation antigens can survive after the cessation of immunosuppression with cyclosporin-A (Cy-A), nerve allografts do not. In an attempt to induce immunological unresponsiveness to nerve allografts, we used grafts containing only minor transplantation antigens and varied the duration of Cy-A therapy from 2 to 12 weeks. Our results demonstrated that nerve allografts survived in rats during Cy-A therapy, but when the drug administration ceased, the allografts were rejected. Other factors besides the degree of histoincompatibility and duration of Cy-A treatment must be involved in determining whether or not unresponsiveness develops to allografts after Cy-A withdrawal. We conclude that nerve allograft immunosuppression generated by Cy-A requires regular administration of the drug.

Localized conformational changes induced in a class I major histocompatibility antigen by the binding of monoclonal antibodies.

We describe two monoclonal antibodies, R3/47 and YR1/1, directed against different epitopes of the expressed rat class I major transplantation antigen RT1Aa, that interact with each other so that the binding of one antibody, YR1/1, is greatly enhanced by the binding of the other. The positive interaction between R3/47 and YR1/1 also occurs when using RT1Aa molecules solubilized from cell membranes in detergent. It is therefore unlikely that the molecular environment of the membrane contributes to the interaction. The ability of R3/47 to modify the YR1/1 determinant on the RT1Aa molecule is mediated without any significant loss of potency by highly purified monomeric Fab fragments. This result suggests that the binding of R3/47 to the RT1Aa molecule alters the YR1/1 determinant by initiating a propagated conformational change in the antigen.

Functional expression of a mouse H-2Kb gene isolated from non-expressing teratocarcinoma cells.

Embryonal carcinoma cells do not express H-2 antigens or beta 2-microglobulin. Recent studies have suggested that the expression of these antigens is likely to be controlled at the level of transcription. To study the precise organization of the corresponding genes and their possible expression in adult mouse cells, we have isolated H-2-related genes from a genomic cosmid library constructed with PCC4-aza-RI from DNA of EC cells. Clones isolated from the library after stringent hybridization with an H-2 cDNA probe were tested for their ability to direct H-2 antigen synthesis after DNA-mediated gene transfer in a fibroblastic L cell. Four clones have been found to code for the major transplantation antigen H-2Kb. Structural analysis showed that these clones contained the same entire H-2Kb gene, identical to the corresponding gene isolated from differentiated C57Bl/10 cells. Furthermore, the present studies showed that this embryonal carcinoma gene was expressed and was functional when transfected into a differentiated cell.

Immunobiological function of trophoblast to circulating maternal IgG antibodies; antibodies to major transplantation antigens.

IgG anti-A and anti-B were isolated from one patient who showed extremely high anti-A and anti-B antibody titers by the indirect antiglobulin test. These IgG were conjugated with horseradish peroxidase (HRP). A human term placenta of blood type B from a type-O mother was incubated in a solution containing these anti-A and anti-B IgG-HRP conjugates.It is indicated that anti-A and anti-B IgG-HRP are endocytosed into the syncytiotrophoblast but these IgG-HRP lose it's character as immunoglobulin promptly after endocytosis from the results obtained, i.e., positive peroxidase reaction was widely observed inside the trophoblast but immunofluorescence carried out by use of rabbit anti-human IgG/FITC was positive only at the apical portion of the syncytiotrophoblast. On the contrary, in the control experiments used of the same placenta of the experiment and non-specific human IgG-HRP, it is verified that non-specific human IgG was transfered across the syncytiotrophoblast into the fetal side without changing its charactor as immunoglobulin.These results might support our hypothesis published in 1976, under the title of “Immunobiological Function of the Syncytiotrophoblast: A New Theory”, 1 in which we explained how the fetus is prevented from circulating maternal IgG class antibodies aimed at fetal antigens, such as HLA and ABO, etc.

Thymus differentiation and T-cell specificity in nu/nu +/+ mouse aggregation chimaeras.

Thymus development and T cell differentiation were studied in mouse chimaeras produced by aggregating pre-implantation embryos of thymus-deficient nude BALB/c (nu/nu) and wild-type C57BL/6 (+/+) mice and vice versa. Chimaeras showed mosaic distribution of skin and coat pigmentation, of hair follicles, of glucosephosphate isomerase within all tested organs and of lymphocytes expressing the different major transplantation antigens (H-2). When tested for their capacity to generate vaccinia virus-specific and self-H-2 specific cytotoxic T cells, all chimaeras of BALB/c (nu/nu) H-2d in equilibrium C57BL/6 (+/+) H-2b type generated T cells of one or both parental origins that were specific for virus and for self-H-2 of the +/+ (H-2b) type only. In contrast, some BALB/c (+/+) H-2d in equilibrium C57BL/6 (nu/nu) H-2b chimaeras generated vaccinia virus-specific cytotoxic T cells specific for either H-2d (+/+) type or for H-2b (nu/nu) type. These asymmetrical results can be interpreted to indicate the following: (i) The +/+ thymus part alone is functional, but because of asymmetrical cross-reactivities of anti-self-H-2 specificities, the observed T cell restriction phenotypes differ. (ii) Both nu/nu and +/+ thymus parts are functional but immune response defects may be exaggerated in such chimaeras producing unexpected non-responsiveness to vaccinia virus linked to H-2d in H-2b (+/+) in equilibrium H-2d (nu/nu).

Class I-like HLA genes map telomeric to the HLA-A2 locus in human cells.

Among the proteins encoded by the major histocompatibility complex (MHC) are the highly polymorphic class I major transplantation antigens, HLA-A and HLA-B in man and H-2K and H-2D in mouse. Class I loci also include the less polymorphic HLA-C in man and H-2L in mouse. Class I antigens are 45,000-molecular weight (Mr) glycosylated membrane proteins associated on the cell surface with apparently nonpolymorphic beta 2-microglobulin. Linked to the murine MHC are genes encoding the class I-like Qa and Tla antigens, which are closely related structurally to H-2K, D and L, as they are also 45,000-Mr cell surface glycoproteins associated with beta 2-microglobulin. Such class I molecules have also been found on the surface of human T lymphocytes and, in one case, shown to be linked to HLA-A. Recombinant DNA techniques have been used to map DNA fragments to the Tla region in mouse. A similar genetic analysis of the HLA complex is hampered by the lack of congeneic strains and by the small number of intra-MHC recombinants in well studied families. One means of overcoming these problems involves a molecular genetic analysis of human lymphoblastoid cell line (LCL) mutants having gamma-ray-induced physical deletions of HLA DNA to map DNA fragments in the MHC. Here we have applied this method to provide evidence that some human class I-like DNA sequences are telomeric to the A2 locus in LCL 721.

B5 and B15 Associated with Progressive Marek's Disease, Rous Sarcoma, and Avian Leukosis Virus-Induced Tumors in Inbred 15I4 Chickens

RPRL 15I4 inbred chickens are nearly histocompatible except for the B5 and B15 haplotypes that determine the major transplantation antigens. B5B5 and B15B15 15I4 chickens were equally and highly susceptible to tumors induced by Marek's disease, Rous sarcoma, or avian leukosis viruses (ALV), suggesting both of these haplotypes may determine a general influence leading to tumor development. The 15I4 birds were highly susceptible to erythroblastosis, and most developed immunological tolerance to the virus after ALV infection.

Exon shuffling: mapping polymorphic determinants on hybrid mouse transplantation antigens.

The mouse major transplantation antigens H-2K, H-2D and H-2L are highly polymorphic cell-surface glycoproteins which may serve as recognition elements in cell-cell interactions. Each antigen possesses a number of alloantigenic determinants defined by antisera of various specificities. Recently, monoclonal antibodies have been produced which redefine and extend our knowledge of these determinants2,3, but structural information has not yet been correlated with the serological definition of the antigens. We have previously reported the molecular cloning of genes for H-2Ld and H-2Dd transplantation antigens from the BALB/c mouse and the expression of these genes in mouse L cells4,5. To localize the serological determinants to discrete regions of the H-2 protein, we have now constructed new H-2 antigen genes by joining together fragments of the H-2Ld and H-2Dd genes. In L cells, these genes direct the synthesis of hybrid H-2 proteins and by using monoclonal antibodies of defined specificities, we have mapped classically defined serological specificities to structurally defined domains of the transplantation antigen protein. We conclude that polymorphic determinants recognized by monoclonal antibodies are located in functionally distinct portions of the protein.

Transplantation antigens: biological function and role as disease determinants

A summary of data is presented which illustrate that major transplantation antigens are named incorrectly for historical reasons. Their biological function is not really to promote rejection of foreign cells and organs but rather to guide cell-cell interactions of cells involved in immunological defense. Immunological recognition of self-transplantation antigens is determined in the thymus and influences the quality of the immune response, and therefore also disease susceptibility.

Primary structural analysis of the Tla region gene products, QA-2 and TL

The Tla region on the murine 17th chromosome encodes at least three gene products (Qa-1, Qa-2 and TL) which are structurally similar to the major transplantation antigens H-2K, -D and -L. In this communication we compare the tryptic peptides of two Tla region-encoded gene products, Qa-2 and TL. The results indicate that TL and Qa-2 may be only modestly related in amino acid sequence and support the previous suggestion that distinct subfamilies of class I gene products with unique biochemical and functional properties may exist.

Allograft and Mixed Lymphocyte Response Studies in 63 × 151 F6 Chickens

The Ea-B genotype defined by hemagglutination was previously shown to influence resistance to tumors induced by Rous sarcoma virus, Marek's disease virus, and RAV-1 virus in 63 × 151 F4–5 chickens. In this study we show that the B-genotype defined by agglutination was also associated with differences in rapidity of skin graft rejection and in responsiveness of mixed lymphocyte cultures. We conclude that F6 chickens differing for Ea-B antigens also differed for the major transplantation antigens and that the potential locus influencing general tumor resistance may conceivably be any one of the tightly linked subloci in the B-haplotype.