Abstract
One-way mixed lymphocyte cultures employing human adenoid or peripheral blood lymphocytes activate lymphotoxin (LT)-secreting cells. Kinetic analysis of lymphocytes stimulated in mixed culture demonstrates that LT is secreted before the onset of DNA synthesis, but that maximum levels of LT secretion are reached simultaneously with maximum levels of DNA synthesis. Although the response of peripheral leukocytes is qualitatively similar to the response of adenoid-derived lymphocytes, unexplained high nonspecific background levels of toxic material(s) obscure early events in the former response. While cytochalasin B reversibly inhibits LT secretion, mitomycin C treated cultures are still capable of LT secretion. The results suggest that a population of cells exists, which does not require DNA synthesis to develop into effector cells. The requirement for DNA synthesis for the maximal development of effector cells may reside in a separate helper cell population as postulated by the two cell model of the mixed lymphocyte reaction.
Highlights
The mixed lymphocyte culture reaction (MLC), observed when genetically incompatible lymphoeytes are cultured together, is thought to be an in vitro correlate of a primary cell-mediated immune response (CMI) with proliferation and the development of cytotoxic effector cells [1,2,3]
The results presented here suggest that in mixed cultures, DNA synthesis and CMI effector function as measured by LT secretion may be separable but related phenomena, which may be associated with different cell populations
The medium used in the mixed lymphocyte cultures consisted of Eagle's minimum essential medium supplemented with 100 U/ml penicillin, 100 t~g/ml streptomycin, 3.2/~g/ml garamycin, 20 t*g/ml amphotericin ]3, 4 mM glutamine, nonessential amino acids, 1 mM sodium pyruvate, 5 × 10-SM mercaptoethanol (EtSH), and 10% pooled A + heat-inactivated human serum (HS), (MEM + 10% HS)
Summary
The mixed lymphocyte culture reaction (MLC), observed when genetically incompatible lymphoeytes are cultured together, is thought to be an in vitro correlate of a primary cell-mediated immune response (CMI) with proliferation and the development of cytotoxic effector cells [1,2,3]. While the relationship between the lymphocytes which synthesize D N A and the cytotoxic cells is not clear, it is generally accepted that DNA synthesis is required for the development of the cytotoxic effector cells [9, 10]. It has been demonstrated in many species [11], including man [12], that the lymphocytes involved in MLC are thymus-derived.
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