Augmentation of proliferation and generation of specific cytotoxic cells in human mixed lymphocyte culture reactions by colchicine

Manikkam Suthanthiran, Kurt H. Stenzel, Albert L. Rubin, Abraham Novogrodsky

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Cellular proliferation and generation of cytotoxic lymphocytes in mixed lymphocyte culture reactions (MLC) results from cellular interactions initiated by individual differences in cell surface structures determined by the HLA-D locus. Cellular microtubular assemblies (MTA) modulate cell shape and surface architecture and have also been implicated in mediating stimulatory signals from the lymphocyte plasma membrane to intracellular sites. To assess the role of MTA in alloactivation, we tested the effect of colchicine, a microtubule-disrupting alkaloid, on the MLC. Diametrically opposite results were observed depending upon the colchicine treatment protocol. Brief exposure of stimulating cells to 10-6 M colchicine resulted in an increase in [3H]thymidine incorporation from 30, 694 ± 2787 to 47,345 ± 4361 cpm/culture (mean ± SEM) (P < 0.001), exposure of responding cells to 10-6 M colchicine resulted in an increase from 33,054 ± 4012 to 46,790 ± 5458 cpm/culture (P < 0.01), and exposure of both cells to 10-6 M colchicine resulted in an increase from 33,054 ± 4012 to 52,685 ± 6720 cpm/culture (P < 0.01). However, direct addition of colchicine to a final concentration of 10-6 M to MLCs at different times resulted in complete suppression of proliferation when added as late as 96 hr, and 63% suppression when added at 120 hr. Pretreatment of stimulating, responding, or both cells with lumicolchicine did not enhance proliferation. Pretreatment of cells with 10-6 and 10-4 M colchicine enhanced proliferation, while pretreatment with 10-2 M colchicine prevented blastogenesis. The potentiation of proliferation induced by colchicine was evident as early as 48 hr after the initiation of the MLC. Generation of specific cytotoxic cells in the MLC was also enhanced by exposing lymphocytes to colchicine prior to the proliferative phase in six out of eight experiments (specific chromium release = 168% of control) despite constant effector:target ratios. These findings indicate that early disruption of microtubules leads to enhanced cellular proliferation and generation of cytotoxic lymphocytes in allogeneic one-way MLCs and suggest that the state of polymerization of the MTA may modulate immune responses involving cell-cell interactions.

Original languageEnglish
Pages (from-to)379-391
Number of pages13
JournalCellular Immunology
Volume50
Issue number2
DOIs
Publication statusPublished - 15 Mar 1980
Externally publishedYes

Fingerprint

Mixed Lymphocyte Culture Test
Colchicine
Lymphocytes
Microtubules
Cell Proliferation
HLA-D Antigens
Cell Shape
Chromium
Lymphocyte Activation
Clinical Protocols
Alkaloids
Individuality
Cell Communication
Polymerization
Thymidine
Cell Membrane

ASJC Scopus subject areas

  • Immunology

Cite this

Augmentation of proliferation and generation of specific cytotoxic cells in human mixed lymphocyte culture reactions by colchicine. / Suthanthiran, Manikkam; Stenzel, Kurt H.; Rubin, Albert L.; Novogrodsky, Abraham.

In: Cellular Immunology, Vol. 50, No. 2, 15.03.1980, p. 379-391.

Research output: Contribution to journalArticle

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abstract = "Cellular proliferation and generation of cytotoxic lymphocytes in mixed lymphocyte culture reactions (MLC) results from cellular interactions initiated by individual differences in cell surface structures determined by the HLA-D locus. Cellular microtubular assemblies (MTA) modulate cell shape and surface architecture and have also been implicated in mediating stimulatory signals from the lymphocyte plasma membrane to intracellular sites. To assess the role of MTA in alloactivation, we tested the effect of colchicine, a microtubule-disrupting alkaloid, on the MLC. Diametrically opposite results were observed depending upon the colchicine treatment protocol. Brief exposure of stimulating cells to 10-6 M colchicine resulted in an increase in [3H]thymidine incorporation from 30, 694 ± 2787 to 47,345 ± 4361 cpm/culture (mean ± SEM) (P < 0.001), exposure of responding cells to 10-6 M colchicine resulted in an increase from 33,054 ± 4012 to 46,790 ± 5458 cpm/culture (P < 0.01), and exposure of both cells to 10-6 M colchicine resulted in an increase from 33,054 ± 4012 to 52,685 ± 6720 cpm/culture (P < 0.01). However, direct addition of colchicine to a final concentration of 10-6 M to MLCs at different times resulted in complete suppression of proliferation when added as late as 96 hr, and 63{\%} suppression when added at 120 hr. Pretreatment of stimulating, responding, or both cells with lumicolchicine did not enhance proliferation. Pretreatment of cells with 10-6 and 10-4 M colchicine enhanced proliferation, while pretreatment with 10-2 M colchicine prevented blastogenesis. The potentiation of proliferation induced by colchicine was evident as early as 48 hr after the initiation of the MLC. Generation of specific cytotoxic cells in the MLC was also enhanced by exposing lymphocytes to colchicine prior to the proliferative phase in six out of eight experiments (specific chromium release = 168{\%} of control) despite constant effector:target ratios. These findings indicate that early disruption of microtubules leads to enhanced cellular proliferation and generation of cytotoxic lymphocytes in allogeneic one-way MLCs and suggest that the state of polymerization of the MTA may modulate immune responses involving cell-cell interactions.",
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