title: A large quantity of CD3-/CD19-/CD16- lymphocytes in malignant pleural effusion from a patient with recurrent cholangio cell carcinoma.
abstract: Tumor infiltrating lymphocytes (TILs) are candidates for adoptive cellular immunotherapy. Here we report on a patient whose TILs presented unusual lymphocyte antigens. Pleural effusions were collected from a 47-year-old man with recurrent cholangio cell carcinoma and malignant effusion. Effusion-associated lymphocytes (EALs) were separated by Ficoll-Hypaque gradient, and the EAL phenotype was determined by flow cytometry. The percentage of positive cells was determined for each lymphocyte-related differentiation antigen. The percentages of CD3+, CD19+, and CD16+ lymphocyte subpopulations among EALs were 20%, 7%, and 3%, respectively. Nearly 70% of EALs were CD3-/CD19-/CD56-/CD16- cells. The phenotypes of peripheral blood lymphocytes (PBLs) collected simultaneously from the patient's peripheral blood were CD3+ (52%), CD19+ (20%), and CD16+ (20%). When EALs were cultured in medium without pleural effusion, T cell-related antigens, but not B cell- or natural killer (NK) cell-related antigens, were newly expressed on EALs, and this expression reached a plateau after 48 h in culture. The proportions of CD3+, CD19+, and CD16+ cells were 69%, 7%, and 3%, respectively. However, when EALs were cultured in medium with pleural effusion, increased expression of T cell-related antigens was not observed; the proportions of CD3+, CD19+, and CD16+ cells were 16%, 6%, and 1%, respectively. Neither total cell numbers nor cellular viability of EALs changed significantly after in-vitro culture, suggesting that significant proliferation or death of EALs did not occur during the culture period. Co-culture of the patient's PBLs with autologous pleural effusion for 96 h did not alter the expression of lymphocyte-related antigens on the PBLs. These results indicate that expression of T cell-related antigens, but not B cell- or NK cell-related antigens, on EALs was blocked temporarily by the malignant pleural effusion. This is the first report concerning the existence of a large quantity of unclassified lymphocytes in which the T cell-related antigens were reversibly masked in the malignant pleural effusion.
authors.affiliation: Department of Cancer Therapy and Research, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Hideya
authors.initials: H
authors.lastname: Onishi
authors.forename: Takashi
authors.initials: T
authors.lastname: Morisaki
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Kuga
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Katano
authors.forename: Fukashi
authors.initials: F
authors.lastname: Doi
authors.forename: Akihiko
authors.initials: A
authors.lastname: Uchiyama
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sugitani
authors.forename: Junji
authors.initials: J
authors.lastname: Wada
authors.forename: Kazuo
authors.initials: K
authors.lastname: Chijiiwa
authors.forename: Masao
authors.initials: M
authors.lastname: Tanaka
mesh_terms: D018941:Antigens, CD19 / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D018281:Cholangiocarcinoma / Q000276:immunology* / Q000628:therapy; D006801:Humans; D016219:Immunotherapy, Adoptive; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000096:biosynthesis; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D009364:Neoplasm Recurrence, Local / Q000276:immunology* / Q000628:therapy; D010844:Picibanil / Q000494:pharmacology; D016066:Pleural Effusion, Malignant / Q000276:immunology* / Q000628:therapy; D017452:Receptors, IgG / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D018941:Antigens, CD19; D017252:CD3 Complex; D017452:Receptors, IgG; D010844:Picibanil; D007371:Interferon-gamma