Human Mixed-Lymphocyte Culture Reaction: Genetics, Specificity, and Biological Implications

Dupont, Bo; Hansen, John A.

doi:10.1016/s0065-2776(08)60320-x

Cited by 126 publications

(36 citation statements)

References 224 publications

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“…Although the suppressive activity detected in these experiments appears to be nongenetically restricted in most cases, the simultaneous presence of an additional, undetected, genetically restricted suppressor is not eliminated. A genetically restricted suppressor cell (31) (23), but the stimulator capacity of group III patients' cells also appeared to depend on the mitomycin C sensitivity of the suppressor cell activity in the population. The suppressor cell activity of group III patient cells appears to be mitomycin sensitive during the 4-to 6-d period and resistant at later periods.…”

Section: Resultsmentioning

confidence: 95%

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Changes in Lymphocyte Activity after Thermal Injury

Miller¹,

Baker²

1979

J. Clin. Invest.

185

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A B S T R A C T The high incidence of fatal septicemia associated with severe thermal injury is believed to result from a loss of immunocompetence. To detect bum-mediated immune defects, lymphocyte function in peripheral blood leukocytes from 18 individuals sustaining 20-80% full thickness thermal burns was investigated. We examined the kinetics of the mitogen responses, the development of suppressive activity, and the correlation of mononuclear cell functional abnormalities with the incidence of sepsis. Patients were divided into three groups corresponding to their clinical course. The phytohemagglutinin responses of Ficoll-Hypaque purified leukocytes from eight ofthese patients (group III) were normal at day 1-2 after injury, but were significantly depressed (mean 16% of normal) at days 5-10 after injury. All of these group III patients experienced multiple, severe, septic episodes, and septic mortality was 75%. The other 10 burned individuals showed either augmented (group II) or unaltered (group I) mitogen responsiveness.Concomitant with evaluation of their mitogen responses, the cells of burn patients were assessed for development of suppressive activity by addition to ongoing normal mixed leukocyte reactions (MLR). Only the addition of mononuclear cells with depressed phytohemagglutinin responsiveness (group III) significantly decreased MLR proliferation (mean 80% reduction) by the previously highly responsive, normal MLR combinations. Addition of cells from group III burn patients collected immediately after injury had no suppressive effect. Addition of cells from patients in group I or II or of normal individual's cells had no suppressive effect. These experimental results strongly suggest that a suppressive mononuclear cell is at least partially responsible for the decreased immunocompetence of bum patients.

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Section: Resultsmentioning

confidence: 95%

“…This poor immune response might have resulted because the normal individual's cells used as stimulators were too genetically similar to the patients' cells (23). Because the patients were not HLA typed, this probably did occur in some cases.…”

Section: Resultsmentioning

confidence: 99%

Changes in Lymphocyte Activity after Thermal Injury

Miller¹,

Baker²

1979

J. Clin. Invest.

185

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“…Histocompatibility typing for the HLA-A, B, and C loci was performed according to the standard National Institutes of Health two-stage microcytotoxicity technique by using eosin dye exclusion method (13). A micromixed lymphocyte culture test was used for mixed lymphocyte culture (MLC) and Dw determination (14). Lymphocytes from individual donors were separated by Ficoll-hypaque method.…”

Section: Methodsmentioning

confidence: 99%

Familial discoid lupus erythematosus associated with heterozygote c2 deficiency

Belin

Bordwell

Einarson

et al. 1980

Arthritis & Rheumatism

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Two siblings with chronic discoid lupus erythematosus and several family members were found with heterozygous C2 deficiency. An association with histocompatibility markers HLA‐B 18 and HLA‐D w2 was demonstrated, and the slow allotype of factor B was present. Linkage studies in this family suggested a close linkage between the C2 deficiency gene and genes coding for B18, Dw2, and BfS antigens. One HLA‐A CB/DBF recombinant was observed showing closer linkage between HLA‐D and Bf than between HLA‐B and Bf.

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“…The stimulation of peripheral blood lymphocytes (PBL) with mitogenic lectins (1,3) and allogeneic cells (2) has become a common laboratory procedure for the clinical monitoring of immunopathological conditions, in histocompatibility reactions before organ transplantation, and in experimental studies on cell-mediated immunity (1,2,5). In the usual assay, lymphocyte blastogenesis is induced in cultures incubated in the wells of microtiter plates and assayed by the incorporation of 3H-thymidine (3H-TdR), followed by DNA precipitation and measurement of the radioactivity in a scintillation counter (4,6).…”

mentioning

confidence: 99%

“…In the usual assay, lymphocyte blastogenesis is induced in cultures incubated in the wells of microtiter plates and assayed by the incorporation of 3H-thymidine (3H-TdR), followed by DNA precipitation and measurement of the radioactivity in a scintillation counter (4,6). However, this procedure requires a relatively long assay, ranging from 6 to 24 h (2,5); it also requires the manipulation of radioactive material and scintillation cocktails containing toxic volatile components and also their disposal in a controlled manner to avoid environmental contamination.…”

mentioning

confidence: 99%

DNA analysis of stimulated lymphocytes by automatic sampling for flow cytometry

1988

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A microsample delivery system (MSDS) was tested for automatic flow cytometry (FCM) analysis of DNA synthesis in stimulated human peripheral blood lymphocytes (PBL) cultivated in wells of microtiter plates. After incubation, either for 1-3 days with phytohemagglutinin, concanavalin A, and pokeweed mitogen, or for 7 days with allogenic PBL, the cells, while in the wells, were washed in hypotonic Tris buffer and stained with ethidium bromide-RNAse solution.The results obtained from quintuplicate replicated wells, each of the five containing the same control or stimulated cultures, were reproducible in terms of the number of nuclei counted in each histogram of control, mitogen-stimulated PBL, and mixed lymphocyte cultures (MLC). Using a computer program that superimposes histograms and calculates their differences on the scale of fluorescence intensity, it was possible to quantify the intensity of the response to the mitogenic stimuli.This approach to the study of lymphocyte proliferation offers not only a simpler and faster analysis of DNA synthesis than the method of 3H-thymidine incorporation, but it also allows for the analysis of other FCM parameters, such as forward and 90" light scatter and double fluorescence labelling of PBL nuclei.

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Human Mixed-Lymphocyte Culture Reaction: Genetics, Specificity, and Biological Implications

Cited by 126 publications

References 224 publications

Changes in Lymphocyte Activity after Thermal Injury

Changes in Lymphocyte Activity after Thermal Injury

Familial discoid lupus erythematosus associated with heterozygote c2 deficiency

DNA analysis of stimulated lymphocytes by automatic sampling for flow cytometry

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