Willard A. Barnes

Generation of tumor-specific cytolytic T lymphocytes from peripheral blood of cervical cancer patients by in vitro stimulation with a synthetic human papillomavirus type 16 E7 epitope.

OBJECTIVEnApproximately 90% of squamous carcinomas of the cervix harbor the human papillomavirus and type 16 has been detected in nearly 50% of cases. Recent studies in mice have shown that the human papillomavirus type 16 E7 oncoprotein contains peptide epitopes that are processed and presented in association with a major histocompatibility antigen for recognition by cytolytic T lymphocytes. We investigated whether an epitope from human papillomavirus type 16 E7 could be used to generate specific human cytolytic T lymphocytes in patients with cervical carcinoma.nnnSTUDY DESIGNnAfter radiation therapy, three patients with antigen HLA-A2 and with locally advanced cervical cancer underwent leukapheresis. Epitope-specific cytolytic T lymphocytes were generated from the peripheral blood mononuclear cells by in vitro stimulation with autologous peripheral blood mononuclear cells pulsed with a human papillomavirus type 16 E7, HLA-A2-restricted, synthetic peptide, E7(11-20) (YMLDLQPETT).nnnRESULTSnIn two patients cytolytic T lymphocytes were capable of E7(11-20)-specific, HLA-A2-restricted cytolysis of the peptide-pulsed, HLA-matched, T2 target cell line. Cytolytic T lymphocytes from one of these patients also demonstrated specific cytolysis against the HLA-A2+, HPV-16+ CaSki cervical cancer cell line but did not lyse either HLA-A2+, HPV-16- MS-751 cells or HLA-A2-, HPV-16- HT-3 cells.nnnCONCLUSIONSnThese experiments demonstrate that novel cytolytic T lymphocytes that recognize a human papillomavirus type 16 E7 epitope can be generated by using the peripheral blood mononuclear cells from irradiated patients with cervical cancer. In addition, because CaSki cells were specifically lysed by the cytolytic T lymphocytes, these data indicate that the peptide E7(11-20) is endogenously processed and presented on the cell surface of the CaSki cells. The demonstration of epitope-specific lysis of cytolytic T lymphocytes of HPV-16+ cervical cancer cells supports further efforts to develop human papillomavirus peptide-based vaccines or antigen-specific adoptive immunotherapy for the prevention and treatment of cervical carcinoma.

p53 polymorphism and risk of cervical cancer

Storey and co-workers have reported data suggesting that individuals homozygous for arginine at residue 72 of p53 (p53Arg) are about seven times more susceptible to invasive cervical cancer than individuals who carry at least one proline at that position (p53Pro). These preliminary data were supported by in vitro evidence demonstrating that the E6 oncoprotein of human papilloma virus (HPV) degrades p53Arg more efficiently than p53Pro. We have now tested specimens from a total of 1,309 women in three studies for p53 polymorphisms. We find that p53Arg is not associated with an increased risk of preinvasive or invasive cervical neoplasia; indeed, there is a tendency for p53Arg to be associated with a decreased risk of neoplasia.

Obesity as a potential risk factor for adenocarcinomas and squamous cell carcinomas of the uterine cervix

Hormonal factors may play a more prominent role in cervical adenocarcinoma than squamous cell carcinoma. The authors evaluated whether obesity, which can influence hormone levels, was associated with adenocarcinoma and squamous cell carcinoma.

Associations between smoking and adenocarcinomas and squamous cell carcinomas of the uterine cervix (United States)

AbstractObjectives: Few studies of smoking and cervical carcinoma have addressed the rare cervical adenocarcinomas or used DNA-based tests to control for human papillomavirus (HPV) infection.nMethods: This multicenter case–control study included 124 adenocarcinoma cases, 307 community controls (matched on age, race, and residence to adenocarcinoma cases), and 139 squamous carcinoma cases (matched on age, diagnosis date, clinic, and disease stage to adenocarcinoma cases). Participants completed risk-factor interviews and volunteered cervical samples for PCR-based HPV testing. Polychotomous logistic regression generated adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for both histologic types.nResults: Eighteen percent of adenocarcinoma cases, 43% of squamous carcinoma cases, and 22% of controls were current smokers. After control for HPV and other questionnaire data, adenocarcinomas were consistently inversely associated with smoking (e.g. current: OR = 0.6, 95% CI 0.3–1.1; ≥1 pack per day: OR = 0.7, 95% CI 0.4–1.3), while squamous carcinomas were positively associated with smoking (e.g. current: OR = 1.6, 95% CI 0.9–2.9; ≥1 pack per day: OR = 1.8, 95% CI 1.0–3.3). Results in analyses restricted to HPV-positive controls were similar.nConclusion: Smoking has opposite associations with cervical adenocarcinomas and squamous carcinomas. Although both histologic types are caused by HPV and arise in the cervix, etiologic co-factors for these tumors may differ.

Comprehensive analysis of human leukocyte antigen class I alleles and cervical neoplasia in 3 epidemiologic studies.

To comprehensively explore the relationship between human leukocyte antigen (HLA) class I alleles and cervical neoplasia, a subset of participants from 3 large US and Costa Rican cervix studies were typed for HLA class I alleles. Study subjects were women with cervical cancer or high-grade squamous epithelial lesions (HSILs; n=365) or low-grade squamous epithelial lesions (LSILs; n=275) or who were cytologically normal (control subjects; n=681). Allele-disease associations were assessed by logistic regression analysis. Consistent associations across all studies were observed for HLA-CW*0202 with a combined odds ratio of 0.53 (95% confidence interval [CI], 0.29-0.89) for cancer or HSILs and 0.58 (95% CI, 0.37-1.04) for LSILs, compared with control subjects and adjusted for study. This finding supports the hypothesis that a single allele may be sufficient to confer protection against cervical neoplasia. Given the relationship between HLA-C and its receptors on natural killer (NK) cells, a role is proposed for NK function in human papillomavirus infection and cervical neoplasia.

Family history as a co-factor for adenocarcinoma and squamous cell carcinoma of the uterine cervix: results from two studies conducted in Costa Rica and the United States.

Previous work suggests that cervical cancer may aggregate in families. We evaluated the association between a family history of gynecological tumors and risk of squamous cell and adenocarcinomas of the cervix in 2 studies conducted in Costa Rica and the United States. The Costa Rican study consisted of 2,073 women (85 diagnosed with CIN3 or cancer, 55 diagnosed with CIN2 and 1,933 controls) selected from a population‐based study of 10,049 women. The U.S. study consisted of 570 women (124 with in situ or invasive adenocarcinomas, 139 with in situ or invasive squamous cell carcinomas of the cervix and 307 community‐based controls) recruited as part of a multicentric case‐control study in the eastern part of the United States. Information on family history of cervical and other cancers among first‐degree relatives was ascertained via questionnaire. Information on other risk factors for cervical cancer was obtained via questionnaire. Human papillomavirus (HPV) exposure was assessed in both studies using broad spectrum HPV L1‐based PCR testing of exfoliated cervicovaginal cells and in Costa Rica by additional testing of plasma collected from participants for antibodies against the L1 protein of HPV types 16, 18, 31 and 45 by ELISA. A family history of cervical cancer in a first‐degree relative was associated with increased risk of squamous tumors in both studies (odds ration [OR] = 3.2 for CIN3/cancer vs. controls; 95% confidence interval [CI] = 1.1–9.4 in Costa Rica; OR = 2.6 for in situ/invasive squamous cell carcinoma cases vs. controls, 95% CI = 1.1–6.4 in the Eastern United States study). These associations were evident regardless of whether the affected relative was a mother, sister or daughter of the study participant. Furthermore, observed effects were not strongly modified by age. In Costa Rica, the effect persisted in analysis restricted to HPV‐exposed individuals (OR = 3.0; 95% CI = 1.0–9.0), whereas in the Eastern United States study there was evidence of attenuation of risk in analysis of squamous carcinoma cases restricted to HPV positive women (OR = 1.4; 95% CI = 0.29–6.6). No significant association was observed between a family history of cervical cancer in a first‐degree relative and adenocarcinomas (OR = 1.3; 95% CI = 0.43–3.9). History of gynecological tumors other than cervical cancer in a first‐degree relative was not significantly associated with risk of disease in either study. These results are consistent with a role of host factors in the pathogenesis of squamous cell cervical cancer, although familial aggregation due to shared environmental exposures cannot be ruled out.

The significance of paraaortic node status in carcinoma of the cervix and endometrium

Survival in 115 women with cervical carcinoma and 55 patients with carcinoma of the endometrium was correlated with the status of paraaortic lymph nodes and the clinical stage of disease. The survival probability at 48 months for patients with cervical cancer who had negative paraaortic lymph nodes was 86% for stage IB in comparison to 67% for those with advanced disease (clinical stages II, III, and IV). The difference of 19% between these two groups was not statistically significant nor was there a statistical difference in survival between early and advanced disease groups when paraaortic nodes contained tumor (37 vs 20%). For all stages of cervical cancer, survival was 80% with negative nodes and 20% with positive nodes at 48 months. In patients with endometrial carcinoma and negative paraaortic lymph nodes, projected survival at 48 months was 72% for stage I and 77% for stages II and III. When paraaortic lymph nodes demonstrated metastatic disease, survival uniformly decreased to 27% of patients with stage I cancer and 25% of those with stage II and III disease. For all stages of endometrial cancer, survival was 76% with negative nodes and 26% with positive nodes at 48 months. The presence or absence of metastatic disease in the paraaortic lymph nodes is an important prognostic factor that appears to be more useful in predicting survival than clinical stage of disease in patients with cervical and endometrial cancer.

Identification of human papillomavirus type 16 in primary and recurrent cervical cancer following radiation therapy

Formalin-fixed, paraffin-embedded tissue blocks from 13 women with cervical carcinoma that recurred following radiation therapy were evaluated for the presence of human papillomavirus (HPV) by in situ hybridization using ribonucleic acid 35S-labeled probes for HPV types 6, 11, 16, and 18. Ten of thirteen patients also had pretreatment biopsies from their primary tumors available for analysis. HPV 16 was detected in both primary and recurrent lesions in 4 women. In 1 case, HPV was detected in the primary tumor and not in the recurrence. HPV 16 was also present in three recurrent cancers from which primary lesions were not available for probing. Radiation therapy did not alter the hybridization signal strength or pattern, suggesting that the HPV genome copy number was not significantly affected. The persistence of HPV 16 in recurrent cervical carcinoma is consistent with the theory that HPV plays a role in maintaining the malignant state.

Case-Control Analysis of Clostridium difficile—Associated Diarrhea on a Gynecologic Oncology Service

Objective: The incidence, morbidity, and risk factors associated with Clostridium difficile-associated diarrhea (CDAD) were studied in a group of gynecologic oncology patients. Methods: A case-control analysis of gynecologic oncology patients with CDAD was carried out from August 1986 through January 1989 in a university medical center. Results: One hundred twenty-three stool samples were tested for C. difficile using the CDT latex agglutination test (Marion Diagnostics, Kansas City, MO). Thirty episodes of CDAD developed in 23 patients. From August 1986 through July 1988, the incidence was stable at 1.5 episodes/100 admissions. From August 1988 through January 1989, the incidence increased to 9.9 episodes/100 admissions (P = 0.005). Compared with patients with nonspecific antibiotic-associated diarrhea, the study patients were hospitalized longer prior to the development of symptoms (mean 15.2 vs. 9.2 days, P = 0.006) and were admitted more frequently with diarrhea (37% vs. 11%, P = 0.015). The rates of surgery, chemotherapy, and radiation therapy were similar. Fever (57% vs. 14%, P < 0.001), abdominal pain (40% vs. 6%, P < 0.001), bloody stools (27% vs. 3%, P = 0.006), and leukocytosis (64% vs. 26%, P = 0.011) were more common among the study cases. The duration, indication, and number of antibiotics administered were similar, though once started, the mean time to symptoms was longer in the study cases (13.7 vs. 6.1 days, P = 0.004). Seven relapses, 1 death, and 1 unplanned colostomy occurred among women with CDAD. Conclusions: C. difficile is a serious cause of nosocomial morbidity in gynecologic oncology patients. Diarrhea developing after antibiotic exposure is more likely to be associated with C. difficile in patients whose symptoms develop several days after completing antibiotics and in patients with a history of CDAD.