Flow cytometric analysis of platelet activation throughout normal gestation.

OBJECTIVE: To measure platelet activation in normal pregnancy, before and after stimulation with agonists, with a whole blood flow cytometric technique. METHODS: In a cross-sectional study, 5 mL of whole blood was collected from healthy volunteers (nine in the first trimester, ten in the second trimester, 35 in the third trimester, and 32 nonpregnant controls). Platelets were treated with an agonist (thrombin or U-46619, a thromboxane A2 analogue) or buffer and were exposed to saturating concentrations of monoclonal antibodies directed against platelet membrane glycoproteins (GPs): 7E3 (fibrinogen receptor GPIIb/IIIa), S12 (alpha granule marker P-selectin), and 6D1 (von Willebrand factor receptor GPIb). Mean fluorescence intensity was determined for 5000 platelets per sample by using a flow cytometer. RESULTS: In the absence of agonist, no significant difference between groups was found in antibody binding. At no stage of pregnancy were circulating activated platelets detected. Platelets from third-trimester subjects bound significantly less 7E3 than platelets of controls or of first- or second-trimester subjects after stimulation with high-dose thrombin (P < .05 for all comparisons). Down-regulation of 6D1 on platelets after stimulation with high-dose U-46619 was significantly greater in third-trimester gravidas than in controls or first-trimester subjects (P < .05). CONCLUSION: Pregnancy does not increase the percentage of activated platelets in the circulation. Platelet reactivity is altered in the third trimester, as evidenced by decreased antibody binding to fibrinogen receptor epitope and enhanced down-regulation of a von Willebrand factor receptor epitope.

Response of spleen lymphocytes shortly after thymectomy in adult mouse.

The blastogenesis response to the phytomitogens, PHA-P, Con A and PWM was used to assess the effect of adult thymectomy on the spleen lymphocytes of C57B1 mice. The mitogenic response to the phytomitogens was determined by 3H-thymidine uptake. The changes produced in theta-antigen bearing spleen lymphocytes were also evaluated making use of theta antibodies from AKR/S mice previously injected with splenic and thymic lymphocytes from CBA/J mice. The present results show that the response to mitogens PHA-P and Con A is reduced early after thymectomy while the response to PWM was only slightly reduced. There was not any correlation between the disminished response to mitogens and the changes observed in theta bearing spleen lymphocytes.