Low-Cost Method to Monitor Patient Adherence to HIV Antiretroviral Therapy Using Multiplex Cathepsin Zymography
- 329 Downloads
Monitoring patient adherence to HIV antiretroviral therapy (ART) by patient survey is inherently error prone, justifying a need for objective, biological measures affordable in low-resource settings where HIV/AIDS epidemic is highest. In preliminary studies conducted in Ethiopia and South Africa, we observed loss of cysteine cathepsin activity in peripheral blood mononuclear cells of HIV-positive patients on ART. We optimized a rapid protocol for multiplex cathepsin zymography to quantify cysteine cathepsins, and prospectively enrolled 350 HIV-positive, ART-naïve adults attending the Themba Lethu Clinic, Johannesburg, South Africa, to test if suppressed cathepsin activity could be a biomarker of ART adherence (103 patients were included in final analysis). Poor adherence was defined as detectable viral load (>400 copies/ml) or simplified medication adherence questionnaire, 4–6 months after ART initiation. 86 % of patients with undetectable viral loads after 6 months were cathepsin negative, and cathepsin-positive patients were twice as likely to have detectable viral loads (RR 2.32 95 % CI 1.26–4.29). Together, this demonstrates proof of concept that multiplex cathepsin zymography may be an inexpensive, objective method to monitor patient adherence to ART. Low cost of this electrophoresis-based assay makes it a prime candidate for implementation in resource-limited settings.
KeywordsMonitoring Zymography Cysteine protease Infectious disease AIDS Sub-Saharan Africa
Human Immunodeficiency Virus
Combination active antiretroviral therapy
Liquid chromatography–tandem mass spectroscopy
Lopinavir boosted with ritonavir
Non-nucleoside reverse transcriptase inhibitors
Nucleoside reverse transcriptase inhibitors
Peripheral blood mononuclear cells
Simplified medication adherence questionnaire
Tenofovir disoproxil fumarate
World Health Organization
This work was completed partially with funding from a Creative and Novel Ideas in HIV Research (CNIHR) grant sponsored by the National Institutes of Health and The University of Alabama at Birmingham Center for AIDS Research (CFAR) program (5P30A1027767) (MOP and DE), and funding from NIH Award Number DP2OD007433 from the Office of the Director, National Institutes of Health (MOP), American Heart Association (RLG), and United States Agency for International Development (USAID) (DE). This study is made possible by the generous support of the American people through Cooperative Agreement AID 674-A-12-00029 from the United States Agency for International Development (USAID). The contents are the responsibility of the authors and do not necessarily reflect the views of USAID or the United States Government and do not necessarily represent the official views of the Office of the Director, National Institutes of Health or the National Institutes of Health. We would like to extend our gratitude to Hazel Molefe and Anna Segoneco from CHRU and Keshendre Moodley and Lindi Coetzee from the University of the Witwatersrand. We would like to thank Hannah Song from University of Toronto for assistance. We would like to acknowledge the directors and staff of Themba Lethu Clinic (TLC), CHRU, HE2RO, and Right to Care (RTC)—a PEPFAR (US President’s Emergency Plan for AIDS Relief)-funded NGO. We would like to acknowledge the Gauteng Provincial and National Department of Health for providing care for the patients at TLC as part of the National Comprehensive Care, Management and Treatment (CCMT) of HIV and AIDS program. Lastly, we would like to sincerely thank the patients attending the Themba Lethu Clinic for their continued trust in the treatment and care provided at the clinic.
MOP and DE conceived the project, conducted experiments, performed analysis, and wrote the paper. PMK, IKP, LMR, AWK, and RLG conducted experiments and performed analysis; LM, DS, WA, and CP helped recruit patient populations and design low-resource considerations to conduct these studies in these environments. All authors have edited and approved final manuscript.
Compliance with Ethical Standards
Conflict of interest
The author(s) declare that they have no competing interests.
- 1.World Health Organization. (2015). Global health observatory (GHO) data. http://www.who.int/gho/hiv/en/.
- 4.Goldman, J. D., Cantrell, R. A., Mulenga, L. B., Tambatamba, B. C., Reid, S. E., Levy, J. W., et al. (2008). Simple adherence assessments to predict virologic failure among HIV-infected adults with discordant immunologic and clinical responses to antiretroviral therapy. AIDS Research and Human Retroviruses, 24, 1031–1035.CrossRefGoogle Scholar
- 8.Grant, R. M., Lama, J. R., Anderson, P. L., McMahan, V., Liu, A. Y., Vargas, L., Goicochea, P., Casapia, M., Guanira-Carranza, J. V., Ramirez-Cardich, M. E., Montoya-Herrera, O., Fernandez, T., Veloso, V. G., Buchbinder, S. P., Chariyalertsak, S., Schechter, M., Bekker, L. G., Mayer, K. H., Kallas, E. G., Amico, K. R., Mulligan, K., Bushman, L. R., Hance, R. J., Ganoza, C., Defechereux, P., Postle, B., Wang, F., McConnell, J. J., Zheng, J. H., Lee, J., Rooney, J. F., Jaffe, H. S., Martinez, A. I., Burns, D. N., Glidden, D. V., & T. iPrEx Study. (2010). Preexposure chemoprophylaxis for HIV prevention in men who have sex with men. The New England Journal of Medicine, 363, 2587–2599.Google Scholar
- 14.de Nooijer, R., Bot, I., von der Thusen, J. H., Leeuwenburgh, M. A., Overkleeft, H. S., Kraaijeveld, A. O., et al. (2009). Leukocyte cathepsin S is a potent regulator of both cell and matrix turnover in advanced atherosclerosis. Arteriosclerosis, Thrombosis, and Vascular Biology, 29, 188–194.CrossRefGoogle Scholar
- 18.Lutgens, E., Lutgens, S. P., Faber, B. C., Heeneman, S., Gijbels, M. M., de Winther, M. P., et al. (2006). Disruption of the cathepsin K gene reduces atherosclerosis progression and induces plaque fibrosis but accelerates macrophage foam cell formation. Circulation, 113, 98–107.CrossRefGoogle Scholar
- 28.Wilson, S., Hashamiyan, S., Clarke, L., Saftig, P., Mort, J., Dejica, V. M., & Bromme, D. (2009). Glycosaminoglycan-mediated loss of cathepsin K collagenolytic activity in MPS I contributes to osteoclast and growth plate abnormalities. American Journal of Pathology, 175, 2053–2062.CrossRefGoogle Scholar
- 32.R. O. S. A. Department of Health (2010). The South African Antiretroviral treatment guidelines 2010, http://www.uj.ac.za/EN/CorporateServices/ioha/Documentation/Documents/ART%20Guideline.pdf.
- 34.Truax, R. E., Powell, M. D., Dietrich, M. A., French, D. D., Ellis, J. A., & Newman, M. J. (1993). Cryopreservation of bovine buffy coat leukocytes for use in immunologic studies. American Journal of Veterinary Research, 54, 862–866.Google Scholar
- 36.South African National Department of Health. (2010). South African National Department of Health Antiretroviral Therapy Guidelines 2010. www.sahivsoc.org/upload/documents/Summary_The_South_African_Antiretroviral_Treatment_2010.pdf.
- 38.Hamers, R. L., Sigaloff, K. C., Wensing, A. M., Wallis, C. L., Kityo, C., Siwale, M. et al. (2012). Patterns of HIV-1 drug resistance after first-line antiretroviral therapy (ART) failure in 6 sub-Saharan African countries: Implications for second-line ART strategies. Clinical Infectious Diseases, 54, 1660–1669.CrossRefGoogle Scholar
- 39.Wallis, C. L., Papathanasopolous, M. A., Fox, M., Conradie, F., Ive, P., Orrell, C., et al. (2012). Low rates of nucleoside reverse transcriptase inhibitor (NRTI) resistance detected in a well monitored cohort in South Africa accessing antiretroviral therapy. Antiviral Therapy, 17, 313.CrossRefGoogle Scholar
- 41.Orrell, C., Harling, G., Lawn, S. D., Kaplan, R., McNally, M., Bekker, L. G., & Wood, R. (2007). Conservation of first-line antiretroviral treatment regimen where therapeutic options are limited. Antivir Ther, 12, 83–88.Google Scholar
- 42.Hoffmann, C. J., Charalambous, S., Sim, J., Ledwaba, J., Schwikkard, G., Chaisson, R. E., et al. (2009). Viremia, resuppression, and time to resistance in human immunodeficiency virus (HIV) subtype C during first-line antiretroviral therapy in South Africa. Clinical Infectious Diseases, 49, 1928–1935.CrossRefGoogle Scholar
- 44.Van Damme, L., Corneli, A., Ahmed, K., Agot, K., Lombaard, J., Kapiga, S., Malahleha, M., Owino, F., Manongi, R., Onyango, J., Temu, L., Monedi, M. C., Mak’Oketch, P., Makanda, M., Reblin, I., Makatu, S. E., Saylor, L., Kiernan, H., Kirkendale, S., Wong, C., Grant, R., Kashuba, A., Nanda, K., Mandala, J., Fransen, K., Deese, J., Crucitti, T., Mastro, T. D., Taylor, D., & F. E.-P. S. Group. (2012). Preexposure prophylaxis for HIV infection among African women. The New England Journal of Medicine, 367, 411–422.Google Scholar