Phenotypic analysis of splenocyte subsets following acute morphine treatment in the rat

Prior studies by our laboratory demonstrated that a single injection of morphine produces dose-dependent, naltrexone-reversible, suppressive effects in assays of mitogen-stimulated lymphocyte proliferation and natural killer (NK) cell cytotoxicity in the spleen. The present study used flow cytometry to assess directly whether acute morphine treatment produces these immune alterations by altering the leukocyte composition of the spleen. In agreement with our previous findings, morphine suppressed the concanavalin A-stimulated proliferation of T cells, lipopolysaccharide-stimulated proliferation of B cells, and NK cell cytotoxicity in the spleen. However, the same morphine treatment protocol did not alter the total number of splenic leukocytes, the percentage of live splenic leukocytes (as assessed by forward-scatter versus side-scatter histograms), or the relative number of CD4(+)CD3(+) T cells, CD8(+)CD3(+) T cells, CD45RA/B(+) B cells, NKR-P1A(hi)CD3(-) NK cells, NKR-P1A(lo)CD3(+) T cells, CD11b/c(+)HIS48(-) monocytes/macrophages, or CD11b/c(+)HIS48(+) granulocytes in the spleen. These findings indicate that the effects of a single sc dose of morphine on functional measures of immune status in the spleen do not result from a redistribution of splenic leukocytes; instead, morphine's effects likely result from direct alterations in leukocyte activities.