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Biomedical subjects

P Iványi

Publications and source records attributed to P Iványi.

At least 19 recordsLinked to original sources

[Identification of HLA-D allele regions using DNA analysis].

The successful bone marrow transplantation presents the highest demands on the compatibility between donor and recipient in the antigen products of HLA system, namely in transplantations performed in unrelated individuals. Traditionally used serotyping gives rather exact results in class I HLA antigen determination, but when typing of class II antigens, at least 25% errors occur. Our papers presents a comparison of different molecular genetics methods used in class II HLA antigen typing. First results of class II HLA typing using RFLP technique, as performed in the Institute of Hematology and Blood Transfusion, are summarized.

Alleles↗

Molecular analysis of a functional subtype of HLA-B27. A possible evolutionary pathway for HLA-B27 polymorphism.

The structure of a new HLA-B27 subtype antigen B27.4(B27D), distinguishable from the HLA-B27.1, B27.2, and B27.3 subtypes by cytolytic T lymphocytes and isoelectric focusing, has been established by comparative peptide mapping and sequence analysis. HLA-B27.4 differs from the main B27.1 subtype in the same two changes of aspartate-77 to serine-77 and valine-152 to glutamate-152, which distinguish the B27.1 and B27.3 subtypes. In addition, there are two other amino acid changes of histidine-114 to aspartate-114 and of aspartate-116 to tyrosine-116, which are unique to B27.4. The close structural relationship between B27.3 and B27.4 explains the similarity of these two subtypes in terms of T cell recognition. The presence of the two single amino acid differences between B27.3 and B27.4 within a span of three residues in the linear sequence provides a new example, suggesting that gene conversion-like mechanisms play a major role in the diversification of HLA-B27. A comparison of the structure of HLA-B27.4 with those of B27.1, B27.2, and B27.3 in the context of their ethnic distribution suggests that the diversification of the HLA-B27 antigens is an ongoing process that has continued after the separation of the major ethnic groups. A tentative evolutionary model for HLA-B27 polymorphism is proposed.

Amino Acid Sequence↗

Polymorphic and autoreactive H-2-specific monoclonal antibody isolated after injections of syngeneic Sendai virus-coated lymphocytes.

An H-2-specific monoclonal antibody (mAb Q-1) was obtained from B10.Q (H-2q) mice injected with syngeneic Sendai virus-coated cells. The IgM monoclonal antibody recognizes the public determinant H-2.25 shared by H-2k (Kk) and H-2r haplotypes and cross-reacts with H-2d, H-2s, H-2p, and H-2q cells, the latter being the haplotype of the challenged B-cell donor. The binding of mAb Q-1 to H-2d, H-2s, H-2q, and H-2p cells was lower than to H-2k and H-2r and of decreasing affinity but could be clearly distinguished from the negative reactions with H-2b and H-2f cells. MAb Q-1 distinguishes between Sendai virus-coated and uncoated lymphocytes only cells with low-affinity binding. On virus-coated or infected (H-2p, H-2q, H-2d, H-2s) cells lysis was stronger than on normal lymphocytes. We interpret the enhanced lysis of Sendai virus-positive cells by mAb Q-1 to be due to recognition of a modified exposure of public H-2 determinants induced by Sendai virus.

Animals↗

Identification of new B27 subtypes (B27C and B27D) prevalent in oriental populations.

With the aid of alloreactive cytotoxic T lymphocytes, three subtypes of HLA-B27 can be defined: B27W, B27K, and B27C (non-B27W and non-B27K). The B27C subtype can be distinguished by 1D-IEF, is not recognized by B27W- or B27K-restricted influenza-A-specific cytotoxic T lymphocytes, and is prevalent in Oriental populations. Preliminary data indicate that the various B27 subtypes (W, K, C) are in different linkage disequilibria with HLA-C antigens. A fourth subtype of B27 (non-B27W and non-B27K), designated as B27D, was detected by 1D-IEF. (See accompanying paper by Neefjes et al.) The finding of four B27 subtypes indicates that the serologically defined B27 antigen comprises a family of various, strongly cross-reactive class-I molecules.

Asian People↗

An improved biochemical method for the analysis of HLA-class I antigens. Definition of new HLA-class I subtypes.

A simple method is described for the biochemical analysis of HLA class I antigens. It is a modification of a previously published procedure for one-dimensional isoelectric focusing (1D-IEF), giving improved resolution and offering larger sample capacity. One million viable cells suffice for analysis, and no more than 25 muCi of radioisotope (35S-methionine) are required. The usefulness of the method is illustrated by the characterization of a total of four biochemically distinct subtypes of HLA-B27, three subtypes of HLA-A24, two subtypes of HLA-A11, three subtypes of HLA-A2, two subtypes of HLA-Bw60, two subtypes of HLA-Bw62, and four subtypes of HLA-B35 in a panel of 24 cells selected for the expression of HLA-B27. We envision that this technique will allow a rigorous classification of HLA-A,B antigens into novel subtypes. Populations of distinct ethnic origin are especially of interest in this regard. This technique might also be used as an additional criterion for the official classification of HLA-A,B antigens.

HLA Antigens↗

Fine specificity of human HLA-B7-specific cytotoxic T-lymphocyte clones. I. Identification of HLA-B7 subtypes and histotopes of the HLA-B7 cross-reacting group.

Three HLA-B7-specific cytotoxic T lymphocyte (CTL) clones are described. Their fine specificity is examined by extensive panel and family studies. Four CTL-defined subtypes of HLA-B7 could be found: HLA-B7.1, -B7.2, -B7.3, and -B7.4. Biochemical evidence was obtained in one-dimension isoelectric focusing for only two subtypes. The estimated subtype frequencies in the Caucasian population are presented. Furthermore, a linkage disequilibrium between HLA-A29 and HLA-B7.3 is suggested. The CTL clones exerted distinct "extra-reactivities", i.e., lysis of some non-B7 target cells. These extra reactivities could be assigned to the cross-reacting group of HLA-B7.

Cells, Cultured↗

Natural autoreactive H-2-specific serum antibodies in a group of BALB/cBy (H-2d) mice.

Natural autoreactive H-2-specific antibodies were detected by lymphocytotoxic test in sera of 15 BALB/cBy (H-2d) mice maintained in the same cage. The antibodies could be detected only for a short period of time; they reacted strongly with H-2d and H-2s and weakly with H-2k,q,r cells. Such antibodies could not be identified in sera of other groups of mice of the same strain maintained in the same environment. All serum-positive mice had a shorter life span than control mice. However, neither a real link between the antibodies and the disease nor a clear understanding of the disease was achieved.

Animals↗

Anti-MHC immunity detected prior to intentional alloimmunization. III. Natural autoreactive H-2-specific antibodies.

Cell fusion was performed between spleen cells from young BALB/cBy (H-2d) mice which have never been immunized and SP2/0 mouse plasmacytoma cells. A monoclonal H-2 specific cytotoxic IgM antibody was obtained (By-1) which detected a new public biregional H-2 specificity, H-2.m210. The mcAb By-1 reacted strongly with H-2Kd, Dd, and H-2s antigens, gave weak cross-reactions with H-2Kk, Dq, H-2r, and H-2v antigens and was negative with H-2b, H-2f, H-2p, and H-2Ld antigens. A polymorphic reaction pattern was also observed on a panel of lymphocytes from B10.W strains. The intriguing finding on this reaction pattern was the reactivity on H-2d cells, including the syngeneic BALB/cBy and truly autologous cells. As shown by capping and immunoprecipitation experiments on H-2d cells and by studies on H-2d-transfected mouse L cells, the target molecules for McAb By-1 were H-2Kd and H-2Dd molecules. The BALB/cBy mouse, from whose spleen cells the McAb By-1 was obtained, survived after the fusion experiment, and serum was examined for the presence of cytotoxic H-2-specific antibodies during the rest of its life. At the time of the fusion, no autoreactive serum antibodies were found, but about 4 months later, we found in the serum of this mouse autoreactive H-2-specific cytotoxic IgM antibodies. The serum antibodies followed the same reaction pattern as that of the McAb By-1. As far as we know, this is the first report of autoreactive H-2-specific antibodies in serum of a mouse which has never been immunized and of the first natural autoreactive H-2-specific monoclonal antibody.

Animals↗

Anti-major histocompatibility complex immunity detected prior to intentional alloimmunization. I. Naturally occurring H-2-specific antibodies in C57BL/KaLwRij (H-2b) mice.

Naturally occurring, H-2-specific, lymphocytotoxic antibodies were detected in 3-10% of young adult and in 10-40% of aged C57BL/KaLwRij (H-2b) mice. The antibodies were of the IgM class and occurred in low titers, but occasionally a high titer was found. The antibodies detected public lymphocyte-membrane antigens controlled by genes identical with, or closely linked to class-I H-2K and H-2D genes. Antibodies against 7 different allogeneic H-2 haplotypes were detected but sera of individual mice exerted different reaction patterns and some specificities occurred more frequently than others. Although the occurrence of the antibodies was age dependent, thymus involution, gammapathies, autoimmunity, the presence of other natural lymphocyte-specific antibodies, and polyclonal or nonspecific stimulation could not be related to the occurrence of natural H-2-specific antibodies. Several possible explanations of natural H-2-specific antibodies exist. We propose that determinants of complex altered self-MHC (MHC + X) antigen(s) triggered the production of H-2-restricted antibodies that recognize H-2-public determinants on normal allogeneic cells.

Age Factors↗

Anti-major histocompatibility complex immunity detected prior to intentional alloimmunization. II. Monoclonal H-2-specific antibodies obtained from an unimmunized C57BL/KaLwRij (H-2b) mouse.

A monoclonal 'natural' anti-H-2 IgM antibody produced by a hybridoma cell line OL-3.17 (H-2 m. 209) is described. The OL-3.17 monoclonal antibody was obtained by hybridization of spleen B cells from an unimmunized C57BL/Ka (H-2b) mouse in the serum of which simultaneously an IgM kappa paraprotein of high concentration and a natural H-2-specific antibody of high titer was detected. The monoclonal antibody OL-3.17 reacted strongly with H-2d and H-2s and weakly with H-2k,q,r lymphocytes, thereby detecting a hitherto unknown H-2 public determinant. The target molecules for OL-3.17 cocapped with class-I H-2 antigens, but immunoprecipitation of H-2 antigens was not achieved. This is the first monoclonal H-2-specific antibody obtained from a mouse without intentional immunization and, with high probability, was derived from a B-cell clone which produced natural H-2-specific antibodies detectable in the serum of the original mouse.

Age Factors↗

A cloned cytotoxic T-lymphocyte (CTL) line recognizing a subtype of HLA B27.

The lymphoblastoid cell-line JY (HLA-A2,2;B7,7;C-; DR4,w6) was used to stimulate T cells from donor HG (HLA-A2,w23; B40,w44; Cw4; DRw6,7). Cloned CTL line were obtained by limiting dilution after tertiary stimulation. Strong cytotoxic activity on stimulator cells was found with all CTL clones obtained. One of the clones (HG-31 recognized a subtype of the HLA-B7 antigen. In this paper, we describe another long-term cloned CTL line (HG-61). This line, when tested on a panel of 107 target cells from unrelated individuals, recognized a subtype of HLA-B27 (B27 "K"). There was no significant association with any other HLA antigen. The cloned CTLs were T8+ and their cytotoxic activity could be blocked by the monoclonal antibody W6/32 which recognizes a framework determinant on HLA-A, -B, and -C molecules. In families, reactivity with cells of the CTL line (HG-61) segregated with HLA. It is concluded that the CTL line interacts with an antigenic determinant shared between the HLA-B7 antigen and JY and the subtype of HLA-B27 (B27 "K"), or detects products of a gene closely linked to HLA-B, not revealed by present-day serology.

Antibodies, Monoclonal↗

Activation of cytotoxic T lymphocytes in HLA-A, -B and -C-identical responder-stimulator pairs. I. Variations in generation of anti-class-II CTL in primary mixed lymphocyte cultures.

Cytotoxic T lymphocytes were activated in primary one-way mixed lymphocyte cultures of cells matched for serologically defined HLA-A, -B and -C antigens. In 16 out of the 29 combinations mismatched for the HLA-D/DR antigens, cell-mediated lympholysis of the stimulator cells occurred. The specificity of 5 selected cytotoxic T lymphocytes was studied in detail. Three of these cytotoxic T lymphocytes recognize antigenic determinants associated with HLA-Bw35 (Breuning et al. 1984, II). The 2 other cytotoxic T lymphocytes failed to lyse T-target cells enriched by rosetting with sheep red blood cells, whereas target cells from the 'non-T' fraction were strongly lysed, indicating that antigenic determinants associated with Class-II HLA molecules were the targets recognized by these cytotoxic T lymphocytes. This notion was supported by a study of a panel of HLA-typed third-party target cells. One cytotoxic T-lymphocyte population preferentially lysed HLA-DR2-positive target cells. Family studies, including a family with a recombination between HLA-B and -D, showed that the target antigen recognized by the latter cytotoxic T lymphocyte segregated with DR2. The second cytotoxic T-lymphocyte population recognized a determinant associated with DRw8. However, in 13 of the 29 HLA-A-, -B- and -C-identical, D/DR-different combinations, cell-mediated lympholysis of stimulator target cells could not be detected, not even on enriched 'non-T' target cells. Thus, after primary mixed lymphocyte culture of HLA-A-, -B- and C-identical, HLA-D/DR-non-identical cells, cytotoxic T lymphocytes directed against sensitizing Class-II molecules can be detected in some combinations, but not in others.

Cell Adhesion↗

Activation of cytotoxic T lymphocytes in HLA-A, -B and -C-identical responder-stimulator pairs. II. New subtypes of HLA-Bw35.

We compared five cytotoxic T lymphocytes raised by primary mixed lymphocyte cultures of HLA-A, -B and -C serologically identical Bw35-positive responder-stimulator combinations. When tested on a panel of third-party target cells, the reactivity pattern of these cytotoxic T lymphocytes allowed the distinction of three subtypes of HLA-Bw35. Cold-target inhibition experiments and analysis of CTL activity at the clonal level showed the existence of subsets of CTLs directed against distinct antigenic determinants associated with HLA-Bw35.

Cells, Cultured↗

Naturally occurring cytotoxic human antibodies recognize H-2-controlled murine lymphocyte antigens.

Human sera contain cytotoxic naturally occurring (CyNa) antibodies which discriminate between lymph node cells from mice differing only at the H-2 complex. Sera from three healthy subjects (normal human sera, NH sera) and one serum from a patient with multiple sclerosis reacted with cells expressing Db, Kd, Kk, and Kp molecules, respectively. However, the following observations suggested that the binding specificity of these CyNa antibodies is to antigens that are distinct from the classical H-2 antigens: (i) the NH sera did not contain cytotoxic anti-HLA antibodies, (ii) redistribution (capping) of H-2 antigens did not induce resistance to lysis for CyNa antibodies, and (iii) individual variation was demonstrated in the expression of the murine lymphocyte antigens detected by the human CyNa antibodies. The reason for this variation appeared to be different for individual NH serum. A maternal effect influenced the expression of the murine lymphocyte antigen detected by one NH serum (anti-H-2b). The differences detected by another NH serum (anti-H-2p) appeared to be inherited, as shown by progeny testing. We hypothesize that the human CyNa antibodies may be directed against antigens controlled or modified by murine viruses (milk borne or endogenous), whose expression is under the influence of the H-2 complex, and that their production might have been stimulated by the products of human genes homologous to murine viruses.

Aged↗

Allo-immune anti-Iak sera of individual mice detect HLA-DR-associated polymorphism on human B cells.

A murine allo-immune A.TH anti-A.TL (anti-Iak) serum pool and sera from individual immunized mice were tested on a large panel of human DR typed B cells, obtained from peripheral blood (PBL-B cells), and human lymphoid tumour cells. The anti-Iak serum pool appeared to be a useful reagent both in cytotoxicity and immunofluorescence for the examination of the presence of class-2 HLA molecules on tumour cells. Only one discrepancy was observed when the pool was tested in parallel with a xeno-anti-human B-cell serum on the cells of 56 lymphoid tumours. Nine sera from individual mice were tested on a panel of 93 PBL-B cells. Some of these sera reacted significantly more weakly with human B cells positive for the DR5 and/or DR7 antigens.

Animals↗

A search for association of HLA antigens with paranoid schizophrenia. A9 appears as a possible marker.

Sixty-two Dutch patients with the diagnosis paranoid schizophrenia (SCH) were HLA-A-, -B- and -C-typed. An increase in the frequencies of A9 (P = 0.02, relative risk 1.8) and B5 (P = 0.04, relative risk 1.9) was found. Although these correlations do not remain significant after correction for the number of antigens tested, both findings confirm other data from the literature, including the first published report from a population in Sweden. From all hitherto published literature data, the combined relative risks for A9 and B5 is significantly increased. These data strongly indicate that the distribution of HLA antigens among SCH patients is different from the control population.

Gene Frequency↗

Specificity of anti-HLA-B27 cytotoxic T lymphocytes.

Sub-types of HLA-B27 were detected by cytotoxic T lymphocytes (CTL) generated between HLA-A, -B- and -C-identical B27-positive individuals. We now report the specificity of six independent CTL's generated by mixed lymphocyte culture (MLC) of HLA-A, -B and -C serologically identical B27-positive responder and stimulator cells. Three CTL's recognize one sub-type, and three the other. The combined reactivity of all CTL's allows unequivocal "typing" of B27-positive cells for the two different sub-types B27K and B27W. The specificity of two CTL's was analysed by cold-target inhibition. The results indicate that (1) no further sub-types of HLA-B27 can be detected by the CTL's raised in these combinations; (2) the majority of the CTL's is directed against the B27 antigens; and (3) "extra reactions" on B27-negative cells are caused by a subset(s) of CTL's recognizing unknown antigens shared between stimulator and target cells. CTL's raised by stimulation of HLA-B27-negative responder cells with B27-positive cells of either sub-type lysed all B27-positive target cells indiscriminately. In cold-target inhibition, however, B27-positive cells, carrying the sub-type of B27 different from that of the stimulator, could not inhibit the lysis of cells bearing the stimulator sub-type of B27. This indicates the activation, in B27-negative responders, of at least two different groups of CTL clones, one directed against shared determinants of HLA-B27, and one against the HLA-B27 sub-type. Heterogeneity of the HLA-B27 antigen may have implications for studies on the well-known association between this antigen and various diseases.

Child↗