Final Report Summary - PAHPV-1 (HPV Pilot Action Indonesia)
Genital infections with high-risk Human papilloma viruses (HPV), in particular HPV type 16, are extremely common and can cause severe diseases of the female genital tract of which worldwide 250 000 women die per year. Healthy subjects, but not patients, predominantly exhibit strong Th1/Th2 type immunity against E2 and E6, suggesting that these responses are associated with protection. Since HPV related diseases are a major problem in developing countries this project performed the current pilot study in order to chart the percentage of individuals at risk for progressive HPV 16 as well as the immune response against this virus.
During the first half year activities were organised to gain awareness, get the legislation for the project in Indonesia as well as in the Netherlands. Screening facilities were organised by the local staff with help of the Leiden department. The second half of the year was used to organise technical necessities and to overcome the main technical problems caused by loss of cell material due to the freezing procedure.
The two local partners integrated the problem of HPV awareness into an already existing STD-education and prevention program in their respective health care regions. At the same time as the start of this EU project a so called 'see and treat' program in the rural areas surrounding the cooperating centres was started with help of financing from a Dutch grant. This program foresees in screening women for HPV related disorders of the lower genital tract. Local staff consisting of doctors, nurses and laboratory staff has been trained in education, diagnosing abnormal cervices by means of aceto-acid application (VIA-test), cytological screening by means of Pap smears and in case of abnormalities treatment of the lesion with cryosurgery. Educational material has been produced and where necessary translated in the local language.
Patient information and informed consent has been translated into the local language in cooperation with the local partners. An NL-delegation introduced the protocol during the kick-off meeting. Furthermore, the technical aspects of preparing and freezing the blood samples were explained. Administrative and organisational measures to overcome several problems were taken.
Because the local partners preferred analysing all the material in the Netherlands instead of in the local situation, the pap-smears and the blood samples were shipped to Leiden.
HPV typing: In summary 19 different HPV types were found in 27 % of the healthy women. In women with HPV related diseases HPV 52 was the most common type in the premalignant lesions whereas HPV 16 had the highest prevalence in the carcinoma group.
Immunological analysis: PBMC was analysed for reactivity against pools of peptides covering the E2 protein (6 pools), E6 protein (4 pools) and E7 protein (2 pools). As a positive control the response to recall antigens and PHA was tested. No antigen serves as negative control. Stimulated PBMC was tested for Interferon-gamma production by ELIspot according to the standard operation procedure LUMC-TIM002_7. A lot of time was used to improve the technique for sample handling. Although the final numbers are small and still rough it seems that they agree with our earlier results in such a way that in women with HPV related disorders no HPV specific T cell response is found.
During the first half year activities were organised to gain awareness, get the legislation for the project in Indonesia as well as in the Netherlands. Screening facilities were organised by the local staff with help of the Leiden department. The second half of the year was used to organise technical necessities and to overcome the main technical problems caused by loss of cell material due to the freezing procedure.
The two local partners integrated the problem of HPV awareness into an already existing STD-education and prevention program in their respective health care regions. At the same time as the start of this EU project a so called 'see and treat' program in the rural areas surrounding the cooperating centres was started with help of financing from a Dutch grant. This program foresees in screening women for HPV related disorders of the lower genital tract. Local staff consisting of doctors, nurses and laboratory staff has been trained in education, diagnosing abnormal cervices by means of aceto-acid application (VIA-test), cytological screening by means of Pap smears and in case of abnormalities treatment of the lesion with cryosurgery. Educational material has been produced and where necessary translated in the local language.
Patient information and informed consent has been translated into the local language in cooperation with the local partners. An NL-delegation introduced the protocol during the kick-off meeting. Furthermore, the technical aspects of preparing and freezing the blood samples were explained. Administrative and organisational measures to overcome several problems were taken.
Because the local partners preferred analysing all the material in the Netherlands instead of in the local situation, the pap-smears and the blood samples were shipped to Leiden.
HPV typing: In summary 19 different HPV types were found in 27 % of the healthy women. In women with HPV related diseases HPV 52 was the most common type in the premalignant lesions whereas HPV 16 had the highest prevalence in the carcinoma group.
Immunological analysis: PBMC was analysed for reactivity against pools of peptides covering the E2 protein (6 pools), E6 protein (4 pools) and E7 protein (2 pools). As a positive control the response to recall antigens and PHA was tested. No antigen serves as negative control. Stimulated PBMC was tested for Interferon-gamma production by ELIspot according to the standard operation procedure LUMC-TIM002_7. A lot of time was used to improve the technique for sample handling. Although the final numbers are small and still rough it seems that they agree with our earlier results in such a way that in women with HPV related disorders no HPV specific T cell response is found.