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N L Reinsmoen

Publications and source records attributed to N L Reinsmoen.

At least 55 records · Page 3Linked to original sources
PubMed1986

Clonal analysis of HLA-DR and -DQ associated determinants: their contribution to Dw specificities.

In order to investigate the distribution of epitopes recognized by T-cell clones directed against HLA class II products, bulk primed cell populations were generated using cells matched for class I determinants but disparate for class II determinants. Cells were cloned by single cell deposition (FACS IV) or limiting dilution (1 cell/3 wells), and assayed for proliferative and cytolytic function with panels of well-characterized cells. All cytolytic clones generated from an anti-DR4/Dw4/DQw3 priming combination or an anti-DR2/Dw2/DQw1 priming combination lysed essentially all targets sharing the same Dw type as the sensitizing cell. In some cases, other targets were also lysed. For instance, some clones were lytic to targets bearing the same DR antigen but another Dw subtype including a few clones lytic to virtually all cells carrying that DR specificity. An occasional target cell expressing a different DR antigen from the sensitizing cell was also lysed by these clones, in some cases to the same extent of lysis seen on the specific target. Monoclonal antibody inhibition studies identified three groups of clones: the DQ directed clones and clones apparently directed at more than one DR product. However, the number of molecules detected for each haplotype remains to be investigated. Our data indicate that determinants detected on both DR and DQ products are associated with the Dw type of the sensitizing cell showing that there is polymorphism recognized by T cells on both DR and DQ that is subtypic to the serologically defined specificities. Thus, it appears that the bulk T-cell response is a composite of individual clones recognizing distinct determinants on these class II molecules. The implications of these findings for studies of HLA restricted recognition are discussed.

Antibodies↗
PubMed1985

A new HLA-DP (SB) specificity (Cp63) defined in Japanese and Caucasian cell populations by cloned PLT cells.

Cp61, Cp62, and Cp63 are HLA-DP (SB) specificities detected by PLT testing using cloned PLT cells derived from Japanese cell donors. It was confirmed, using Japanese and Caucasian cell panels, that Cp61 is identical to DPw5 and Cp62 is identical to DPw4. Cp63, however, was found not to correlate with any known HLA-DP specificities. The gene frequency of Cp63 was found to be 3.6% in Japanese (N = 42) and 1.9% in Caucasian (N = 52) cell panels of limited size. No triplet assignment of DPw specificity including Cp63 was found in the Japanese and Caucasian cell populations. The relationship between the Cp63 and the HLA antigens demonstrated no significant correlation with HLA-A,B,C,DR, or MT antigens in this study. Linkage between Cp63 and HLA was confirmed from family segregation studies. This is the first report of a new DP specificity termed Cp63.

Asian People↗
PubMed1985

Analysis of human class II antigens by cloned cytolytic T cell reagents: a study using HLA loss mutant lymphoblastoid cell lines and monoclonal antibodies detecting the HLA-DP product(s).

We have utilized cloned T cell reagents and ionizing radiation-induced mutants of an HLA heterozygous lymphoblastoid cell line (LCL) to investigate the determinants detected by the cell-mediated lympholysis (CML) assay. Cells of an LCL clone, 721.501, an HLA haplotype loss mutant expressing the HLA-A2-Cw1-Bw51-DR1-Dw1-DQw1-DPw2-GLO haplotype were used as sensitizing cells for responder cells in vitro. "Cloned" reagents were generated by single-cell deposition of cells of a bulk reagent primed against 721.501 cells. Those clones were screened for cytolytic activity against HLA loss mutant targets (derived from LCL 721) of four different categories; HLA-A2 loss only, A2-Cw1-Bw51 loss, A2-Cw1-Bw51-DR1-DQw1 loss, and the entire HLA haplotype loss. Of 196 clones tested, 36 were cytolytic, including three anti-A2, five anti-Bw51/Cw1, 12 anti-DR1/DQw1, 13 anti-DP region associated with DPw2, and three of undetermined specificity, based on cytolytic patterns against the HLA loss mutant targets. Of 25 anti-HLA class II lytic clones, 23 (92%) fitted the characteristics of helper cell-independent cytolytic T cells (HITc), whereas only two of eight (25%) anti-class I clones were HITc. The 13 anti-DP region clones were divided into three subgroups defined by blocking by anti-FA and not Tü39 monoclonal antibodies (MoAb), by Tü39 and not anti-FA, and by both MoAbs.

Antibodies, Monoclonal↗
PubMed1984

A study of HLA-DR2 associated HLA-Dw/LD specificities.

HLA-Dw2 and Dw12 are both associated with HLA-DR2; however, these specificities accounts for only 86% (161/188) of the DR2+ haplotypes in our North American Caucasian panel. In an attempt to identify new DR2 associated antigenic clusters, we have generated four primed lymphocyte (LD) typing (PLT) reagents in haploidentical familial combination against DR2+ Dw blank haplotypes. These reagents were positively restimulated by 11 of 16 DR2+ Dw blank cells tested, with good discrimination from Dw2 and Dw12+ cells, thus identifying a new antigenic cluster provisionally termed LD-MN2. We have compared the LD-MN2 specificity with the specificity LD-5a defined by two DR2+ HTCs, BAS and REM, (Layrisse, Caracas) which have been included in the pre-1984 Workshop Cluster DB9. Although none of our DR2+ cells gave typing responses to these two HTCs defining LD-5a, PLT studies did indicate an interrelationship between these specificities and with the specificity tb24 defined with the HTC, FJO (Betuel). The LD-5a HTCs, four LD-5a heterozygous cells, and two additional HTCs (WJR-Hansen, Seattle and FJO/tb24--Betuel, Lyon) significantly restimulated the anti-MN2 PLT reagents, though usually not as strongly as the MN2+ cells. MN2+ cells primed against the LD-5a HTCs were restimulated by only the LD-5a+ cells. Dw2+ cells primed against FJO were restimulated by some, but not all MN2+ cells. These results suggest that MN2, tb24, and LD-5a share some determinants, not shared with most cells which type as Dw2 and Dw12, though differing by other stimulatory determinants. These studies emphasize the necessity of studying new antigenic clusters by both PLT and HTC methodologies as well as testing different ethnic groups.

Epitopes↗
PubMed1984

Detection of HLA restricted anti-minor histocompatibility antigen(s) reactive cells from skin GVHD lesions.

Recipients of marrow grafts from HLA-identical, (including MLC non-reactive) sibling donors have a 40-50% incidence of acute graft vs. host disease (GVHD); the involvement of non-HLA linked, minor histocompatibility antigens (miHA) has been implicated. One of the target tissues of GVHD is the skin where morphological analysis of biopsy specimens is often used to support this diagnosis. We have obtained skin biopsy specimens from the site of GVHD lesions, grown the cells in the presence of T cell growth factor (TCGF) and feeder cells, and tested these cultured cells in the primed lymphocyte test (PLT) and cell mediated lympholysis (CML) assay. Five of six cell cultures tested demonstrated secondary proliferative but not cytolytic reactivity; cells from one culture demonstrated both reactivities. The cell culture populations generated are presumably directed against non-HLA antigens, i.e., miHA whose expression is restricted by an HLA antigen of the recipient. The data are consistent with the suggestion that in the majority of these cultures, the restriction element may be a determinant encoded within the HLA-D region: DQ, DR, and possibly DP based on panel testing. Although the number of cases is small, these preliminary data demonstrate the feasibility of this type of culturing system and also suggest that the cellular immunological events leading to the manifestation of skin GVHD lesions may be predominantly a delayed type hypersensitivity reaction.

Adult↗
PubMed1984

Cellular basis of anti-SB response.

Cloning of cells allosensitized in vitro against SB1, SB2, and SB3 antigens was performed by micromanipulation. One hundred and twenty-six clones were tested for both proliferative and cytolytic responses; 14 proliferative, noncytotoxic clones and one clone which demonstrated both proliferative and cytotoxic reactivity specific for SB antigens were obtained. The proliferative noncytotoxic clones tested and the clone with both cytotoxic and proliferative activity were all able to produce IL-2-like activity upon specific antigen stimulation in vitro and were positive for OKT3, OKT4 but negative for OKT8. The proliferative clones fit the characteristics of helper T cell (Th) clones while the clone with both cytotoxic and proliferative reactivities is analogous to the class of antigen-driven, helper cell-independent cytotoxic (HITc) clones. No cytolytic nonproliferative SB specific clones were detected. The prevalent induction of Th clones strongly suggests that the biological function of SB antigens is similar to other class II antigens of HLA. The existence of an SB specific HITc clone demonstrates that a determinant on an SB molecule can induce both proliferative and cytolytic responses.

Antibodies, Monoclonal↗
PubMed1982

Five HLA-D clusters associated with HLA-DR4.

In order to investigate the HLA-D clusters associated with DR4, 54 DR4-positive, Dw4- and Dw10-negative responders, together with selected Dw4- or Dw10-positive responders, were tested with 22 HTCs that define DR4-associated D specificities. The results are consistent with previous data defining four distinct D clusters--Dw4, Dw10, DB3, and DYT--and have identified a new cluster provisionally termed LD40. In addition, the DB3 cluster is complex and appears to give typing response patterns overlapping those of the KT2 cluster originally defined as being associated with DR4 in Japanese populations. Of 116 DR4-positive haplotypes tested, 44% typed as Dw4, 18% were LD40, 16% were Dw10, 9% were DB3, 3% were DYT, and 10% gave no typing response to the HTCs defining any of these clusters. These studies are informative not only in defining the DR4-associated D clusters and in supporting the concept that D and DR cannot be considered identical but also in emphasizing the complexity of the D region.

Asian↗
PubMed1982

Cloned cellular reagents to define antigens encoded between HLA-DR and glyoxylase.

Primed lymphocyte typing reagents have been used to define antigens encoded by genes of a locus (loci) mapping between HLA-DR and glyoxalase I. This locus, which we shall refer to as the third locus of the HLA-D region, has been variously referred to as D beta, PL beta, PL3, and SB. Generating discriminatory primed lymphocyte typing reagents which can be used to define these antigens, however, has been extremely difficult. Donors of responding and stimulating cells for the priming combinations have usually been matched not only for the DR, D, and MB/MT antigens but also for the HLA-A, -B, and -C antigens. Even under these very restricted conditions, not all bulk primed lymphocyte typing reagents that are generated are discriminatory enough to be useful for antigen definition. We have derived "clones" from bulk priming combinations in which stimulator and responder differed for known antigens of this third locus. Even though the bulk reagents that were prepared did not provide discriminatory results, approximately 7-12% of the clones derived from the bulk priming combination proved to be highly discriminatory. We have been able to obtain these results with regard to all three antigens of the third locus so far evaluated. The very ease of screening clones and deriving discriminatory reagents, as compared with screening responder-stimulator combinations, allows the ready derivation of cellular reagents that define the antigens of this third locus.

Antibody Formation↗
PubMed1980

Complexity of the HLA-D region studied by primed-lymphocyte test.

The results obtained by mixed lymphocyte culture (MLC), HLA-D and -DR typing, and primed lymphocyte test (PLT) in the F. family give some indication of the complexity of the HLA-D region in man. Two siblings identical for HLA-A, -B, -C, -D, and -DR by typing are mutually MLC-stimulatory. PLT studies indicate that one of these siblings expresses D-region determinants of both of the mother's haplo-types, suggesting an intra-D recombinant. These results suggest that the D region contains genes for a number of different determinants.

Epitopes↗
PubMed1979

Anomalous mixed lymphocyte culture reactivity between HLA--A, --B, --C, --DR identical siblings.

Complete HLA typing including HLA--A, --B, --C, --DR (D related B cell typing), --D, mixed lymphocyte culture (MLC), and primed lymphocyte testing (PLT), together with complete red blood cell (RBC), glyoxalase (GLO), GBG (Factor B), and phosphoglucomutase 3 (PGM3) typings were performed on a informative family. The five siblings inherited the four possible combinations of parental HLA haplotypes, and two of the siblings were HLA--A, --B, --C and --DR identical. Repeated MLC testing of the family revealed positive mixed lymphocyte reactivity in all combinations. B cell typing for the DR specificities demonstrated no variation from the expected inheritance pattern and specifically no recombination event. GBG and GLO typings militated against a recombination involving the paternal chromosome. HLA--D testing revealed that only one of the HLA--A, --B, --C, and --DR identical siblings gave typing responses to the HLA--Dw3 specificity present on that maternal haplotype. Utilizing HLA haploidentical combinations, lymphocytes were primed against the four parental haplotypes and the non-Dw3 haplotype of interest (Aw24--B8--DRw3--LDY) in the PLT. The sibling inheriting this haplo-type did not restimulate cells primed against the A2--B40--DRW6--LDY specificity. Furthermore, no discrimination was observed in the restimulation of lymphocytes primed against this haplo-type. Possible interpretations of these family data include: a spontaneous mutation, non-major histocompatibility locus (MHC) stimulation, and HLA--DR/D recombination.

B-Lymphocytes↗
PubMed1978

Antigens associated with acute leukemia detected in the primed lymphocyte test.

Mixed lymphocyte culture (MLC) studies of families with several leukemia patients, all potential bone marrow transplant recipients, demonstrated that cells from acute myelogenous leukemia patients (5 of 5) and acute undifferentiated leukemia patients (1 of 4) in relapse stimulated autologous lymphocytes as well as lymphocytes from siblings known to be identical at the major histocompatibility linkage group. In the patients studied, the blast transformation induced by leukemia cells was not detectable when the patient was in remission. Stimulation by leukemia cells also elicited increased responses of the lymphocytes from normal haploidentical siblings, parents, and unrelated individuals as compared to stimulation by normal allogeneic cells or leukemia cells of patients with leukemia in remission. The primed lymphocyte test (PLT) was used successfully to establish HLA-D identity of the leukemia patients and their respective HLA-identical siblings, despite high percentages of circulatory blasts. Utilizing lymphocytes from normal siblings primed against the leukocytes from an HLA-identical sibling with leukemia, we also presented results of PLT's which suggested that the stimulation induced by leukemia cells in MLC was produced by leukemia-associated antigens.

Acute Disease↗