Identification of HLA-DRB1 and HLA-DQB1 identical individuals by a cytokine-based mixed lymphocyte culture.
Cytokine determination in MLC is under discussion as providing more sensitive and specific information regarding host-graft compatibility, and is therefore suggested to represent a new method for transplantation medicine. Little is known, however, about the stimulatory influence of HLA class II antigens and minor lymphocyte-stimulating antigens (Mls). Our results demonstrate that cytokine determination in MLC is suitable to detect identical alleles of HLA-DRB1 and HLA-DQB1. Among more than 100 random MLC experiments, we observed one cytokine pattern similar to the cytokine release detected in a control MLC of HLA-identical siblings, which showed marginal or no secretion of IL-2, sIL-2R, IFN-gamma, TNF-alpha, and IL-6. HLA-typing of these two nonreactive individuals elevated identical HLA-DRB1 and HLA-DQB1 regions, while they differed in the HLA-DP locus. This suggests that HLA-DP has no stimulatory influence on cytokine release. Further investigation of the stimulatory capacity of HLA-DR and DQ showed that HLA-DR is more effective in inducing IFN-gamma release than HLA-DQ. To evaluate the stimulatory influence of human Mls, i.e., human endogenous retroviruses (HERV), we analyzed HERV sequences of nonreactive individuals. Both individuals showed identical HERV patterns. A third individual, who had shown distinct cytokine release in MLC with both nonreactive individuals, differed in the HERV fragments. In conclusion, cytokine determination in MLC is a new method of evaluating the biological relevance of stimulatory antigens after allogeneic stimulation detecting all individual diversities in one experiment.
Danzer
S G
SG
Institute of Immunology and Transfusion Medicine, University of Lübeck School of Medicine, Germany.
A Campo
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C
Kunze
B
B
Kirchner
H
H
Rink
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L
D000483:Alleles; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D016207:Cytokines / Q000378:metabolism*; D006683:HLA-DQ Antigens / Q000235:genetics / Q000276:immunology*; D059866:HLA-DQ beta-Chains; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology*; D059811:HLA-DRB1 Chains; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000378:metabolism; D007519:Isoantigens / Q000235:genetics / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed / Q000379:methods*; D016747:Minor Lymphocyte Stimulatory Antigens / Q000235:genetics / Q000276:immunology; D012190:Retroviridae / Q000235:genetics / Q000276:immunology
D000956:Antigens, Viral; D016207:Cytokines; D006683:HLA-DQ Antigens; D059866:HLA-DQ beta-Chains; C069051:HLA-DQB1 antigen; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; D007519:Isoantigens; D016747:Minor Lymphocyte Stimulatory Antigens; D007371:Interferon-gamma