title: Differential expression of CD3zeta message and protein in tumor infiltrating lymphocytes from solid tumor specimens and malignant ascites from patients with ovarian carcinoma.
abstract: The expression of the CD3zeta subunit was investigated in fresh (uncultured) tumor-infiltrating lymphocytes (TILs) isolated from either solid tumor (ST) specimens or ascites (ASC) from patients with epithelial ovarian carcinoma (EOC). Western blot analysis of CD3zeta immunoprecipitates using anti-CD3zeta rabbit serum revealed that in 6 out of 6 patients with EOC, the CD3zeta protein was absent from ST-TILs. Immunoprecipitation with anti-phosphotyrosine monoclonal antibody (anti-PY20) from ST-TILs from one patient revealed bands co-migrating with the phosphorylated CD3zeta. CD3zeta protein was found to be expressed in only 1 out of 7 ST-TILs from patients with EOC. ASC-TILs were available in 5 of these patients and immunoprecipitation/Western blotting experiments using anti-CD3zeta rabbit serum revealed that CD3zeta protein was expressed in all 5. In addition, CD3zeta protein was expressed in 3 additional ASC-TIL specimens for which ST-TILs were not available. Therefore, the CD3zeta protein was expressed in ASC-TIL isolated from 8 out of 8 patients with EOC. CD3zeta protein was also expressed on peripheral blood mononuclear cells (PBMCs) from patients with EOC and from normal donors. RT-PCR studies of fresh ST-TIL specimens, using CD3zeta-specific primers, revealed that CD3zeta transcripts were absent from 13 out of 21 patients with EOC, down-regulated in 4 patients and present at levels comparable to those found in PBMCs in 4 other patients. In contrast, CD3delta transcripts were present at comparable levels in all specimens. Treatment with recombinant interleukin-2 (rIL-2) (600 IU/ml) restored the expression of CD3zeta protein and transcripts in cultured ST-TILs, whereas fresh ST-TILs did not express CD3zeta, in contrast to fresh ASC-TILs. These results demonstrate differential expression of CD3zeta in ST-TILs versus ASC-TILs in patients with EOC. CD3zeta transcripts and protein were found to be absent from most ST-TILs from patients with EOC, whereas they were expressed in ASC-TILs and PBMCs from such patients.
authors.affiliation: Department of Microbiology and Immunology, Temple University School of Medicine, Philadelphia, PA, USA.
authors.forename: John
authors.initials: J
authors.lastname: Pappas
authors.forename: Alexey D
authors.initials: AD
authors.lastname: Wolfson
authors.forename: Weon J
authors.initials: WJ
authors.lastname: Jung
authors.forename: Emilia L
authors.initials: EL
authors.lastname: Oleszak
authors.forename: C William
authors.initials: CW
authors.lastname: Helm
authors.forename: Ralph S
authors.initials: RS
authors.lastname: Freedman
authors.forename: Alexander Y
authors.initials: AY
authors.lastname: Tsygankov
authors.forename: Chris D
authors.initials: CD
authors.lastname: Platsoucas
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001201:Ascites / Q000378:metabolism / Q000473:pathology; D017252:CD3 Complex / Q000096:biosynthesis*; D015536:Down-Regulation; D005260:Female; D006801:Humans; D047468:Immunoprecipitation; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000097:blood / Q000276:immunology* / Q000473:pathology; D010766:Phosphorylation; D014443:Tyrosine / Q000378:metabolism
chemical_list: D017252:CD3 Complex; C121704:CD3 antigen, zeta chain; D014443:Tyrosine