An approach to the immunophenotypic features of circulating CD4⁺NKG2D⁺ T cells in invasive cervical carcinoma.
BACKGROUND
NKG2D, an activating immunoreceptor, is primarily restricted to NK cells and CD8(+) T cells. The existence of an atypical cytotoxic CD4(+)NKG2D(+) T cell population has also been found in patients with autoimmune dysfunctions. Nonetheless, contradictory evidence has categorized this population with a regulatory rather than cytotoxic role in other situations. These confounding data have led to the proposal that two distinct CD4(+)NKG2D(+) T cell subsets might exist. The immune response elicited in cervical cancer has been characterized by apparent contradictions concerning the role that T cells, in particular T-helper cells, might be playing in the control of the tumor growth. Interestingly, we recently reported a substantial increase in the frequency of CD4(+)NKG2D(+) T cells in patients with cervical intraepithelial neoplasia grade-1. However, whether this particular population is also found in patients with more advanced cervical lesions or whether they express a distinctive phenotype remains still to be clarified. In this urgent study, we focused our attention on the immunophenotypic characterization of CD4(+)NKG2D(+) T cells in patients with well-established cervical carcinoma and revealed the existence of at least two separate CD4(+)NKG2D(+) T cell subsets defined by the co-expression or absence of CD28.


RESULTS
Patients with diagnosis of invasive cervical carcinoma were enrolled in the study. A group of healthy individuals was also included. Multicolor flow cytometry was used for exploration of TCR alpha/beta, CD28, CD158b, CD45RO, HLA-DR, CD161, and CD107a. A Luminex-based cytokine kit was used to quantify the levels of pro- and anti-inflammatory cytokines. We found an increased percentage of CD4(+)NKG2D(+) T cells in patients with cervical cancer when compared with controls. Accordingly with an increase of CD4(+)NKG2D(+) T cells, we found decreased CD28 expression. The activating or degranulation markers HLA-DR, CD161, and CD107a were heterogeneously expressed. The levels of IL-1beta, IL-2, TNF-alpha, and IL-10 were negatively correlated with the percentages of CD4(+)NKG2D(+) T cells in patients with cervical carcinoma.


CONCLUSIONS
Taken together, our results reveal the existence of two separate CD4(+)NKG2D(+) T cell subsets defined by the co-expression or absence of CD28, the latter more likely to be present in patients with cervical cancer.
Departamento de Fisiología, Laboratorio de Inmunología, CUCS, Universidad de Guadalajara, Sierra Mojada # 950, Guadalajara, Jalisco, México. maye_999@hotmail.com.
Mariel
M
Garcia-Chagollan
División de Inmunología, CIBO, Instituto Mexicano del Seguro Social, Guadalajara, Jalisco, México. lfjave@yahoo.com.
Luis Felipe
LF
Jave-Suarez
Departamento de Biología Celular y Molecular, Laboratorio de Inmunobiología, CUCBA, Universidad de Guadalajara, Guadalajara, Jalisco, México. jharamati@gmail.com.
Jesse
J
Haramati
Departamento de Biología Molecular y Genómica, Instituto de Enfermedades Crónico-Degenerativas, CUCS, Universidad de Guadalajara, Guadalajara, Jalisco, México. bueno@cencar.udg.mx.
Miriam Ruth
MR
Bueno-Topete
División de Inmunología, CIBO, Instituto Mexicano del Seguro Social, Guadalajara, Jalisco, México. adry.aguilar.lemarroy@gmail.com.
Adriana
A
Aguilar-Lemarroy
Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Unidad de Biotecnología Médica y Farmacéutica, Guadalajara, Jalisco, México. cestrada.ciatej@gmail.com.
Ciro
C
Estrada-Chavez
Departamento de Biología Molecular y Genómica, Instituto de Enfermedades Crónico-Degenerativas, CUCS, Universidad de Guadalajara, Guadalajara, Jalisco, México. bastidas@cencar.udg.mx.
Blanca Estela
BE
Bastidas-Ramirez
Departamento de Fisiología, Laboratorio de Inmunología, CUCS, Universidad de Guadalajara, Sierra Mojada # 950, Guadalajara, Jalisco, México. analauraps@hotmail.com.
Ana Laura
AL
Pereira-Suarez
Departamento de Fisiología, Laboratorio de Inmunología, CUCS, Universidad de Guadalajara, Sierra Mojada # 950, Guadalajara, Jalisco, México. susana@cucs.udg.mx.
Susana
S
Del Toro-Arreola
D015703:Antigens, CD / Q000097:blood / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D016207:Cytokines / Q000097:blood / Q000276:immunology; D005260:Female; D006801:Humans; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000097:blood / Q000276:immunology*; D009361:Neoplasm Invasiveness; D002583:Uterine Cervical Neoplasms / Q000097:blood / Q000276:immunology* / Q000473:pathology
D015703:Antigens, CD; D016207:Cytokines; C527085:KLRK1 protein, human; D055655:NK Cell Lectin-Like Receptor Subfamily K