Cervical carcinoma in Kampala, Uganda, and the relationship with human immunodeficiency virus and human papillomavirus infections

Introduction: Cervical carcinoma is the commonest tumour in Ugandan women. However, very little is known in the Ugandan population about HPV types associated with invasive cervical cancer, the relative risk of cervical cancer associated with different HPV types, and whether the risk of cervical cancer associated with HPV infection is modified by HIV infection. Furthermore, the nature of HPV negative invasive cervical cancer, especially adenocarcinoma, is yet to be described and studied to find if the use of different types of cervical samples can influence the studies results. Objectives: To study HPV types associated with invasive cervical cancer. To estimate the relative risk for infections with different HPV types, and whether this is modified by HIV infection. To describe the nature of HPV negative cancer, especially adenocarcinoma, and study if the use of different invasive cervical cancer samples can influence the studies results. Subjects and Methods: We conducted studies using 186 archival cases of cervical carcinoma diagnosed between 1967 and 1980, 316 new incident cases of cervical cancer and exfoliated cervical cells from 314 control women from Uganda between September 2004 and December 2006. HPV testing was done using the SPF10 LiPA technology. In paper I, we assessed the prevalence and HPV types. For Article II, we used different antibodies to clarify the nature of HPV negative adenocarcinomas. In Article III, we estimated the odds ratio (OR) of cervical cancer for different HPV infections as well as for HIV infection. In Article IV, we compared HPV detection in formalin-fixed paraffin embedded (FFPE) and fresh samples. Results: In Article I, Out of the 186 cases of confirmed invasive cervical cancer in the study paraffin blocks, 114 were positive for HPV DNA. Specific HPV genotypes were identifiable in 109 cases: HPV 16, 18, 31, 35, 39, 44, 45, 51, 52 and 70. These occurred as single infections in 105 cases (96.3%) and as multiple infections in 4 cases (3.7%). HPV 16 or 18 accounted for 80% (84/105) of cases with a single infection. In Article II, Among the 13/25 HPV negative ADC samples, five were positive for CEA, suggesting endocervical origin, and three were vimentin positive (one had a mucinous endocervical histological pattern and two were ADC, not otherwise specified, most likely of endometrial origin). In Article III, we found a high relative risk of both squamous cell carcinoma and adenocarcinoma of the cervix with HPV 16 and 18, and moderate relative risk with HPV 45. Concurrent infection with HIV did not increase the relative risk with these three HPV types, but was associated with a statistically borderline increased relative risk of squamous cervical carcinoma only with low-risk types and multiple infections with HPV types. Article IV showed comparable HPV detection in both fresh and FFPE tissue by the SPF10 LiPA method, with complete agreement for 59.7%, partial agreement 5.3% and complete disagreement 35.0%. Conclusion: Most cervical carcinomas in Uganda are associated with high-risk HPV types, and concurrent infection with HIV has little impact. Both archival and fresh samples are suitable for HPV detection, and the use of antibodies may be useful in the characterization of HPV negative cervical adenocarcinomas

HPV types, HIV and invasive cervical carcinoma risk in Kampala, Uganda: a case-control study

<p>Abstract</p> <p>Background</p> <p>While the association of human papillomavirus (HPV) with cervical cancer is well established, the influence of HIV on the risk of this disease in sub-Saharan Africa remains unclear. To assess the risk of invasive cervical carcinoma (ICC) associated with HIV and HPV types, a hospital-based case-control study was performed between September 2004 and December 2006 in Kampala, Uganda. Incident cases of histologically-confirmed ICC (N=316) and control women (N=314), who were visitors or care-takers of ICC cases in the hospital, were recruited. Blood samples were obtained for HIV serology and CD4 count, as well as cervical samples for HPV testing. HPV DNA detection and genotyping was performed using the SPF₁₀/DEIA/LiPA₂₅ technique which detects all mucosal HPV types by DEIA and identifies 25 HPV genotypes by LiPA version 1. Samples that tested positive but could not be genotyped were designated HPVX. Odds ratios (OR) and 95% confidence intervals (CI) were calculated by logistic regression, adjusting for possible confounding factors. </p> <p>Results</p> <p>For both squamous cell carcinoma (SCC) and adenocarcinoma of the cervix, statistically significantly increased ORs were found among women infected with HPV, in particular single HPV infections, infections with HPV16-related types and high-risk HPV types, in particular HPV16, 18 and 45. For other HPV types the ORs for both SCC and adenocarcinoma were not statistically significantly elevated. HIV infection and CD4 count were not associated with SCC or adenocarcinoma risk in our study population. Among women infected with high-risk HPV types, no association between HIV and SCC emerged. However, an inverse association with adenocarcinoma was observed, while decrease in CD4 count was not associated with ICC risk.</p> <p>Conclusions</p> <p>The ORs for SCC and adenocarcinoma were increased in women infected with HPV, in particular single HPV infections, infections with HPV16- and 18-related types, and high-risk HPV types, specifically HPV16, 18 and 45. HIV infection and CD4 count were not associated with SCC or adenocarcinoma risk, but among women infected with high-risk HPV types there was an inverse association between HIV infection and adenocarcinoma risk. These results suggest that HIV and CD4 count may have no role in the progression of cervical cancer.</p

The usefulness of immunohistochemistry in tissue microarrays of Human Papillomavirus negative adenocarcinoma of the uterine cervix

Background: The origin of adenocarcinomas presenting on the cervix uteri may be doubtful, i.e. whether it is of cervical or endometrial origin, due to the overlapping morphological features. In HPV negative samples, further tests may be needed to ascertain the nature of the tumours. We aimed to explore the use of immunohistochemistry profiles in tissue microarrays in archived samples of adenocarcinoma (ADC) of the cervix from Uganda that tested negative for HPV DNA. Findings: Five commercially available antibodies were tested in tissue array sections immunostained utilizing the avidin-biotin (AB) technique. In 26 ADC samples, HPV was detected in 13, p16 in 15 (8 in HPV positive and 7 in HPV negative), CEA in 12, vimentin in 6, ER in 0, and PR in 2. Among the 13/25 HPV negative ADC samples, five were positive for CEA suggesting endocervical origin, and three were vimentin positive (one had a mucinous endocervical histological pattern and two were ADC, not otherwise specified, most likely of endometrial origin). Conclusions: The immunoprofiles of ADC with the antibodies studied are rather nonspecific. By using immunohistochemistry in 13 HPV negative ADC, endocervical tumour origin was suspected in five CEA positive cases while two out of three vimentin positive samples were probably of endometrial origin, suggesting that CEA and vimentin may be valuable in distinguishing HPV negative cervical adenocarcinomas from endometrial adenocarcinomas

Determination of the frequency and distribution of APC, PIK3CA, and SMAD4 gene mutations in Ugandan patients with colorectal cancer

Uganda is a developing low-income country with a low incidence of colorectal cancer, which is steadily increasing. Ugandan colorectal cancer (CRC) patients are young and present with advanced-stage disease. In our population, there is a scarcity of genetic oncological studies, therefore, we investigated the mutational status of CRC tissues, focusing in particular on the adenomatous polyposis coli (APC), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), and SMAD4 genes. Our objective was to determine whether there were any differences between other populations and Ugandan patients. We performed next-generation sequencing on the extracted DNA from formalin-fixed paraffin-embedded adenocarcinoma samples from 127 patients (mean (SD) age: 54.9 (16.0) years; male:female sex ratio: 1.2:1). Most tumours were located in the rectum 56 (44.1%), 14 (11%) tumours were high grade, and 96 (75.6%) were moderate grade CRC. Stage III + IV CRC tumours were found in 109 (85.8%) patients. We identified 48 variants of APC, including 9 novel APC mutations that were all pathogenic or deleterious. For PIK3CA, we found 19 variants, of which 9 were deleterious or pathogenic. Four PIK3CA novel pathogenic or deleterious variants were included (c.1397C > G, c.2399_2400insA, c.2621G > C, c.2632C > G). Three SMAD4 variants were reported, including two pathogenic or deleterious variants (c.1268G > T, c.556dupC) and one tolerant (c.563A > C) variant. One novel SMAD4 deleterious mutation (c.1268G > T) was reported. In conclusion, we provide clinicopathological information and new genetic variation data pertinent to CRC in Uganda

Reduced plasma concentrations of vitamin B6 and increased plasma concentrations of the neurotoxin 3-hydroxykynurenine are associated with nodding syndrome: a case control study in Gulu and Amuru districts, Northern Uganda

Introduction: Nodding syndrome was first reported in Uganda in 2003 among internally displaced populations. Risk factors for the syndrome remain unknown. We therefore explored vitamin B6 deficiency and resulting high 3-hydroxykynurenine (3-HK) levels as risk factor for nodding syndrome in Northern Uganda. Methods: Case-control study conducted in Gulu and Amuru districts. Cases were children/young adults with nodding syndrome. Healthy children/young adults were recruited as controls from same community as cases. Data on socio-demographic and other risk factors was collected using questionnaires. Whole blood was collected in EDTA tubes for assay of 3-HK and vitamin B6 using sandwich ELISA. Conditional logistic regression model was used to assess associations. Results: 66 cases and 73 controls were studied. Factors associated with nodding syndrome were being positive for 3-HK (AOR=4.50, p=0.013), vitamin B6 concentration below mean (AOR=7.22, P=0.001), child being taken care of by mother only (AOR=5.43, p=0.011), child being taken care of by guardian (AOR=5.90, p=0.019) and child consuming relief food at weaning (AOR=4.05, p=0.021). Conclusion: Having low vitamin B6 concentration which leads to a build up of 3-hydroxykynurenine concentration in cases as a main risk factor. Therefore, cases should be treated with vitamin B6 and community members should be sensitise to ensure adequate dietary intake of vitamin B6 so that the risk of nodding syndrome among children is averted. We encourage future prospective intervention study to be conducted to assess the effect of low vitamin B6 on the development of nodding syndrome via raised 3-HK concentration.Pan African Medical Journal 2016; 2

Comparison of human papillomavirus detection between freshly frozen tissue and paraffin embedded tissue of invasive cervical cancer

Background: Human Papillomavirus (HPV) detection results comparing paraffin embedded cervical tissue and other cervical specimens have been done with varying degrees of agreement. However, studies comparing freshly frozen specimens and paraffin embedded specimens of invasive cervical carcinomas are lacking. The aim of the study was to compare HPV detection using SPF10 broad-spectrum primers PCR followed by DEIA and genotyping by LiPA25 (version 1) between freshly frozen cervical tissue samples and paraffin embedded blocks of cervical tissue from the same patient. There were 171 pairs of paraffin embedded and freshly frozen samples analyzed from cervical carcinoma cases from Kampala, Uganda. Results: 88.9% (95% CI: 83.2%-93.2%) of paraffin embedded samples were HPV positive compared with 90.1% (95% CI: 84.6%-94.1%) of freshly frozen samples, giving an overall agreement in HPV detection between fresh tissue and paraffin embedded tissue at 86.0% (95% CI: 79.8%-90.8%). Although the proportion of HPV positive cases in freshly frozen tissue was higher than those in paraffin blocks, the difference was not statistically significant (p > 0.05). In both types of tissues, single HPV infections were predominant, with HPV16 accounting for 47% of positive cases. Comparison in the overall agreement, taking into accounts not only positivity in general, but also HPV types, showed a 65% agreement (complete agreement of 59.7%, partial agreement of 5.3%) and complete disagreement of 35.0%. HPV detection in squamous cell carcinomas (SCC) and adenocarcinomas (ADC) was similar in fresh tissue or paraffin blocks (p ≥ 0.05). p16 immunostaining in samples that had at least one HPV negative results showed that 24 out of 25 cases had an over-expressed pattern. Conclusions: HPV DNA detection was lower among ADC as compared to SCC. However, such differences were minimized when additional p16 testing was added, suggesting that the technical issues may largely explain the HPV negative cases

Human Papillomavirus type distribution in invasive cervical cancer in Uganda

<p>Abstract</p> <p>Background</p> <p>We conducted a study aiming to describe Human Papillomavirus (HPV) type distribution in invasive cervical carcinoma in Uganda.</p> <p>Methods</p> <p>191 archival cervical carcinoma samples diagnosed in the Department of Pathology, Makerere University in Kampala between 1968 and 1992 were analysed using a sensitive PCR-Reverse Hybridization Line Probe Assay.</p> <p>Results</p> <p>Out of the 186 cases of confirmed invasive cervical cancer in the study paraffin blocks, 114 were positive for HPV DNA. Specific HPV genotypes were identifiable in 109 cases: HPV 16, 18, 31, 35, 39, 44, 45, 51, 52 and 70. These occurred as single infections in 105 cases (96.3%) and as multiple infections in 4 cases (3.7%). HPV 16 or 18 accounted for 80% (84/105) of cases with single infection.</p> <p>Conclusion</p> <p>The results of this study confirm the role of HPV 16 and 18 in cervical cancer pathogenesis in the Ugandan population. The results suggest that the currently available HPV vaccines against HPV 16 and 18 could possibly prevent the majority of invasive cervical cancers in Uganda.</p

Papillary squamous cell carcinoma of the cervix in Uganda: A report of 20 cases

BACKGROUND: Non-glandular papillary carcinoma of the cervix are uncommon tumours. In Uganda where cervical carcinoma is very common, no cases of papillary squamous cell carcinoma of the cervix has been reported. OBJECTIVES: To ascertain the occurrence and describe the clinicopathological features of papillary squamous cell carcinoma of the cervix in Uganda. STUDY DESIGN: Retrospective review of histologically diagnosed cases of squamous cell carcinoma of cervix with papillary structures. METHODS: Retrospective review of cases of cervical carcinoma diagnosed in the Pathology Department, Makerere University from 1968 to 1973 was done. Cases with features of squamous differentiation and forming papillary pattern were then selected. RESULTS: Twenty cases were encountered and the ages of the patients ranged from 22 to 70 years (mean 46.6 years). Histologically, the tumours had thin to broad fibrovascular cores covered by multilayered squamous epithelium. In five cases, there were areas with very delicate fibrovascular cores covered by monolayered epithelial cells. CONCLUSION: The results of this study show that in Uganda, papillary squamous cell carcinoma of the cervix does occur and is predominantly a disease of older women. The results also confirm that papillary squamous cell carcinoma is a distinct subtype with some variants, and support the hypothesis that squamous cell carcinoma of the cervix is heterogeneous group of tumuors

Papillary squamous cell carcinoma of the cervix in Uganda: a report of 20 cases

Background: Non-glandular papillary carcinoma of the cervix are uncommon tumours. In Uganda where cervical carcinoma is very common, no cases of papillary squamous cell carcinoma of the cervix has been reported. Objectives: To ascertain the occurrence and describe the clinicopathological features of papillary squamous cell carcinoma of the cervix in Uganda. Study Design: Retrospective review of histologically diagnosed cases of squamous cell carcinoma of cervix with papillary structures. Methods: Retrospective review of cases of cervical carcinoma diagnosed in the Pathology Department, Makerere University from 1968 to 1973 was done. Cases with features of squamous differentiation and forming papillary pattern were then selected. Results:Twenty cases were encountered and the ages of the patients ranged from 22 to 70 years (mean 46.6 years). Histologically, the tumours had thin to broad fibrovascular cores covered by multilayered squamous epithelium. In five cases, there were areas with very delicate fibrovascular cores covered by monolayered epithelial cells. Conclusion: The results of this study show that in Uganda, papillary squamous cell carcinoma of the cervix does occur and is predominantly a disease of older women. The results also confirm that papillary squamous cell carcinoma is a distinct subtype with some variants, and support the hypothesis that squamous cell carcinoma of the cervix is heterogeneous group of tumuors