ISOLATION AND PURIFICATION OF AN EARLY-PREGNANCY FACTOR-LIKE MOLECULEFROM CULTURE SUPERNATANTS OBTAINED FROM LYMPHOCYTES OF PREGNANT-WOMEN

Purpose: Our purpose was to determine whether lymphocytes synthesize p roteins during pregnancy, to observe whether one of the proteins synth esized has early pregnancy factor (EPF)-like activity and to isolate a nd purify this molecule from culture supernatants obtained from stimul ated lymphocytes of pregnant women. Methods: Lymphocyte proliferation assay and S-35-methionine labeling were done to study de novo synthesi s of proteins followed by autoradiography to confirm synthesis of prot eins. The rosette inhibition assay was used for detection of the EPF-l ike molecule. Gelfiltration on Sephadex G-100 and RPHPLC were used for purification of the EPF-like molecule. Results: The rate of incorpora tion of S-35-methionine was significantly higher in the lymphocytes of pregnant women compared to those of the control, and autoradiography confirmed the synthesis of proteins during pregnancy. There is a total protein enhancement trend observed during the first trimester that de clines toward term. The EPF-like molecule is observed to be synthesize d during all the trimesters of pregnancy. This molecule, when purified , showed a single homogeneous biologically active peak. Conclusions: T he results indicated that there is an enhancement of existing protein or synthesis of new proteins during pregnancy. The EPF-like molecule i s one of the many proteins synthesized and secreted by lymphocytes dur ing pregnancy chat, when purified, is biologically active.