title: Isolation and purification of an early pregnancy factor-like molecule from culture supernatants obtained from lymphocytes of pregnant women.
abstract: OBJECTIVE
  Our purpose was to determine whether lymphocytes synthesize proteins during pregnancy, to observe whether one of the proteins synthesized has early pregnancy factor (EPF)-like activity and to isolate and purify this molecule from culture supernatants obtained from stimulated lymphocytes of pregnant women.
  
  
  METHODS
  Lymphocyte proliferation assay and 35S-methionine labeling were done to study de novo synthesis of proteins followed by autoradiography to confirm synthesis of proteins. The rosette inhibition assay was used for detection of the EPF-like molecule. Gel filtration on Sephadex G-100 and RPHPLC were used for purification of the EPF-like molecule.
  
  
  RESULTS
  The rate of incorporation of 35S-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 declines toward term. The EPF-like molecule is observed to be synthesized during all the trimesters of pregnancy. This molecule, when purified, showed a single homogeneous biologically active peak.
  
  
  CONCLUSIONS
  The results indicated that there is an enhancement of existing protein or synthesis of new proteins during pregnancy. The EPF-like molecule is one of the many proteins synthesized and secreted by lymphocytes during pregnancy that, when purified, is biologically active.
authors.affiliation: Institute for Research in Reproduction (ICMR), Parel, Mumbai, India.
authors.forename: C
authors.initials: C
authors.lastname: Aranha
authors.forename: U
authors.initials: U
authors.lastname: Natraj
authors.forename: K S
authors.initials: KS
authors.lastname: Iyer
authors.forename: S
authors.initials: S
authors.lastname: Shahani
mesh_terms: D001345:Autoradiography; D018835:Chaperonin 10; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003208:Concanavalin A; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007553:Isotope Labeling; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D008297:Male; D010455:Peptides / Q000302:isolation & purification* / Q000378:metabolism; D011247:Pregnancy / Q000276:immunology*; D011257:Pregnancy Proteins; D011264:Pregnancy Trimesters / Q000276:immunology; D012397:Rosette Formation; D013491:Suppressor Factors, Immunologic / Q000096:biosynthesis / Q000302:isolation & purification*
chemical_list: D018835:Chaperonin 10; D010455:Peptides; D011257:Pregnancy Proteins; D013491:Suppressor Factors, Immunologic; C025124:early pregnancy factor; D003208:Concanavalin A