Visual Inspection of Acetic Acid (VIA) as a Promising Standard for Cervical Cancer Screening

Objective: To evaluate the "false negative" of VIA in our study population compared to HPV DNA test as the reference test or gold standard. Method: We processed the cervical swab from 1,279 patients with negative VIA and detected the HPV DNA by using INNO-Lipa HPV DNA test. Result: From 1,279 women with negative VIA, 65 samples were excluded because of incomplete data and duplicate examination. From the remaining 1,214 women with negative VIA, 39 samples were confirmed to be positive for HPV DNA by both PCR and hybridization, leading to a "false negative" result of 3.21%. Conclusion: This study shows VIA as a very effective method for cervical cancer screening. VIA gives an excellent result, particularly for ectocervix, with minimal cost. Therefore, it is very suitable to be used as cervical cancer screening in developing countries like Indonesia. Keywords: cervical cancer, HPV DNA, negative VIA, screening, VI

Prevalence of High-Risk Human Papillomavirus (HPV) among Negative Visual Inspection of Acetic Acid (VIA)

Objective: Persistence of high-risk HPV infection is known to be the major cause of cervical cancer. It is important to differentiate the genotype of HPV infection, whether it is high, intermediate or low risk. The aim of this study was to assess the prevalence of high-risk HPV types among Indonesian women with negative VIA. Method: We analyzed cervical swabs from 1,214 patients with negative VIA. By using INNO-Lipa HPV DNA test, we detected the HPV DNA and its genotype. Result: From the 1,214 women with negative VIA, 48 (3.95%) samples were confirmed to have positive HPV DNA by using PCR and electrophoresis. However, hybridization test were not able to detect HPV genotypes in 9 samples. These 9 samples were tested again with PCR and electrophoresis and resulted in negative HPV DNA. Among the remaining 39 samples (3.21%), we detected 19 types of HPV, consisting of 13 types of high-risk HPV, 5 types of low-risk HPV, and 1 type of unknown HPV (type X). Conclusion: Among patients with negative VIA, 3.21% was found to be positive for HPV DNA. From this percentage, the prevalence of high-risk HPV is higher than the low-risk and unknown HPV. Therefore we cannot ignore results of negative VIA, particularly in highrisk group, because there is a slight possibility that presence of HPV can be identified, especially the high risk ones which have a tendency to be persistent. We support the importance of HPV DNA test as cervical cancer screening method. Keywords: cervical cancer, high-risk HPV, negative VI

Human Papillomavirus Genotypes and its Prevalence in Normal Population

Objective: Over 200 types of human papillomavirus (HPV) have been recognized on the basis of DNA sequence. Multiple infection is more prone to be persistent than single infection. The purpose of this study is to assess the variation of HPV types and their prevalence among negative VIA as normal population in Indonesian women. Method: We processed cervical swabs from 1,214 patients with negative VIA. HPV DNA and its genotypes were detected using PCR based INNO-Lipa HPV DNA test. We also classified whether each infection is single or multiple. Result: From 1,214 women with negative VIA, 39 (3.21%) samples were positive for HPV DNA. Among them, we detected 19 types of HPV, consisting of 13 types of high-risk HPV, 5 types of low-risk HPV, and 1 type of unknown HPV (type X). The most prevalent type was HPV type 52 (18.31%), followed by type 39 and X with the same proportion (9.86%), and HPV type 16, 18, and 74 (each 8.45%). Of the total 39 HPV-positive samples, 17 (43.6%) showed multiple-type infection and 22 (56.4%) showed single-type infection. The majority of single infection involves high-risk-HPV. The remaining were type 6, 44, 18, 51 and 66, with each single-type infection showing a prevalence of 4.54%. Conclusion: Our study shows that single HPV infection among the negative VIA population are dominated by high-risk type HPV (types 52, 39, 16, and 18). Single infection was more often encountered than multiple infection. Keywords: HPV DNA, HPV genotypes, multiple infection, negative VIA, single infectio

Induction of lymphokine-activated killer activity in rat splenocyte cultures: The importance of 2-mercaptoethanol and indomethacin

The role of 2-mercaptoethanol and indomethacin in the induction of lymphokine-activated killer (LAK) activity by interleukin-2 (IL-2) in rat splenocyte cultures was investigated. Spleens from 4-month-old male rats of five different strains were tested. Splenocytes were cultured for 3-5 days in the presence of IL-2 (1000 U/ml) and LAK activity was assessed by 4-h51Cr release assays with P815 and YAC-1 cells as targets. LAK activity could be induced by IL-2 in splenocytes from all rat strains, but only when 2-mercaptoethanol was present in the culture medium. Optimal LAK activity was induced when the 2-mercaptoethanol concentration in splenocyte cultures was at least 5 μM. Different rat strains showed differences in levels of in vitro induction of LAK activity. In the presence of 2-mercaptoethanol the level of LAK activity induced by IL-2 was high in BN and Lewis rats, intermediate in Wistar and Wag rats, and low in DZB rats. In the absence of 2-mercaptoethanol no or minimal LAK activity was induced. Furthermore we observed that addition of 50 μm indomethacin to the culture medium in the presence of 2-mercaptoethanol augmented the induction of LAK activity to some extent. In the absence of 2-mercaptoethanol, addition of indomethacin resulted only in low levels or no induction of LAK activity. We conclude that for optimal induction of LAK activity by IL-2 in rat splenocyte cultures 2-mercaptoethanol is essential, while indomethacin can only marginally further improve this induction

Association of antigen processing machinery and HLA class I defects with clinicopathological outcome in cervical carcinoma

HLA class I loss is a significant mechanism of immune evasion by cervical carcinoma, interfering with the development of immunotherapies and cancer vaccines. We report the systematic investigation of HLA class I and antigen processing machinery component expression and association with clinical outcome. A tissue microarray containing carcinoma lesions from 109 cervical carcinoma patients was stained for HLA class I heavy chains, β2-microglobulin, LMP2, LMP7, LMP10, TAP1, TAP2, ERAP1, tapasin, calreticulin, calnexin and ERp57. A novel staining evaluation method was used to ensure optimal accuracy and reliability of expression data, which were correlated with known clinicopathological parameters. Partial HLA class I loss was significantly associated with decreased 5-years overall survival (61% vs. 83% for normal expression; P < 0.05) and was associated with decreased 5-years disease-free survival (DFS) (65% vs. 82% for normal expression; P = 0.05). All APM components except LMP10, calnexin and calreticulin were down-regulated in a substantial number of cases and, except ERAP1, correlated significantly with HLA class I down-regulation. LMP7, TAP1 and ERAP1 loss was significantly associated with decreased overall and (except LMP7) DFS (P < 0.05 and 0.005, respectively). ERAP1 down-regulation was an independent predictor for worse overall and DFS in multivariate analysis (HR 3.08; P < 0.05 and HR 2.84; P < 0.05, respectively). HLA class I and APM component down-regulation occur frequently in cervical carcinoma, while peptide repertoire alterations due to ERAP1 loss are a major contributing factor to tumour progression and mortality

High-resolution analysis of HLA class I alterations in colorectal cancer

BACKGROUND: Previous studies indicate that alterations in Human Leukocyte Antigen (HLA) class I expression are frequent in colorectal tumors. This would suggest serious limitations for immunotherapy-based strategies involving T-cell recognition. Distinct patterns of HLA surface expression might conceal different immune escape mechanisms employed by the tumors and are worth further study. METHOD: We applied four-color multiparameter flow cytometry (FCM), using a large panel of alloantigen-specific anti-HLA-A and -B monoclonal antibodies, to study membranous expression of individual HLA alleles in freshly isolated colorectal cancer cell suspensions from 21 patients. RESULTS: Alterations in HLA class I phenotype were observed in 8 (38%) of the 21 tumors and comprised loss of a single A or B alleles in 4 cases, and loss of all four A and B alleles in the other 4 cases. Seven of these 8 tumors were located on the right side of the colon, and those showing loss of both HLA-A and -B membranous expression were all of the MSI-H phenotype. CONCLUSION: FCM allows the discrimination of complex phenotypes related to the expression of HLA class I. The different patterns of HLA class I expression might underlie different tumor behavior and influence the success rate of immunotherapy