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Low-Cost Method to Monitor Patient Adherence to HIV Antiretroviral Therapy Using Multiplex Cathepsin Zymography

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Abstract

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.

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Abbreviations

3TC:

Lamivudine

ART:

Antiretroviral therapy

ARV:

Antiretrovirals

CI:

Confidence interval

EFV:

Efavirenz

HIV:

Human Immunodeficiency Virus

cART:

Combination active antiretroviral therapy

LC/MS:

Liquid chromatography–tandem mass spectroscopy

LPV/r:

Lopinavir boosted with ritonavir

NNRTI:

Non-nucleoside reverse transcriptase inhibitors

NRTI:

Nucleoside reverse transcriptase inhibitors

NVP:

Nevirapine

PBMC:

Peripheral blood mononuclear cells

PI:

Protease inhibitors

PrEP:

Pre-exposure prophylaxis

RR:

Risk ratio

SMAQ:

Simplified medication adherence questionnaire

TDF:

Tenofovir disoproxil fumarate

WHO:

World Health Organization

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Acknowledgments

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.

Authors’ contributions

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.

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    Authors and Affiliations

    1. Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, 315 Ferst Drive, IBB 1308, Atlanta, GA, 30332, USA

      Manu O. Platt, Philip M. Keegan, LaDeidra M. Roberts & Rudolph L. Gleason Jr.

    2. The Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA, USA

      Manu O. Platt, Philip M. Keegan, Ivana K. Parker, LaDeidra M. Roberts, Alexander W. Caulk & Rudolph L. Gleason Jr.

    3. Health Economics and Epidemiology Research Office, Department of Internal Medicine, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa

      Denise Evans

    4. Clinical HIV Research Unit, Department of Internal Medicine, School of Clinical Medicine, University of the Witwatersrand, Johannesburg, South Africa

      Lynne McNamara

    5. The George W. Woodruff School of Mechanical Engineering, Atlanta, GA, USA

      Ivana K. Parker, Alexander W. Caulk & Rudolph L. Gleason Jr.

    6. Department of Biochemistry, Addis Ababa University, Addis Ababa, Ethiopia

      Daniel Seifu

    7. Department of Internal Medicine, Addis Ababa University, Addis Ababa, Ethiopia

      Wondwossen Amogne

    8. Oncology Division, Department of Internal Medicine, University of the Witwatersrand, Johannesburg, South Africa

      Clement Penny

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    1. Manu O. Platt
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    Manu O. Platt and Denise Evans have contributed equally to this work.

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    Platt, M.O., Evans, D., Keegan, P.M. et al. Low-Cost Method to Monitor Patient Adherence to HIV Antiretroviral Therapy Using Multiplex Cathepsin Zymography. Mol Biotechnol 58, 56–64 (2016). https://doi.org/10.1007/s12033-015-9903-0

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