Table of Contents  
ORIGINAL ARTICLE
Year : 2021  |  Volume : 23  |  Issue : 1  |  Page : 46-51

HIV-1 plasma RNA viral load and CD4 cell count in drug-naïve HIV-1 infected patients in Kaduna State, Nigeria


1 Department of Medical laboratory Science, Ahmadu Bello University, Zaria, Nigeria
2 Department of Microbiology, Ahmadu Bello University, Zaria, Nigeria
3 Department of Medical Microbiology, Ahmadu Bello University, Zaria, Nigeria

Date of Submission25-Jul-2019
Date of Decision06-Oct-2019
Date of Acceptance18-Jun-2020
Date of Web Publication28-Apr-2021

Correspondence Address:
Mohammed Ibrahim Tahir
Department of Medical laboratory Science, Ahmadu Bello University, Zaria
Nigeria
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jomt.jomt_27_19

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  Abstract 


Background: The recent guidelines for HIV treatment initiation in Nigeria do not depend on CD4 cell count or plasma viral load however, assessment of the baseline immunologic and virologic markers could indicate prognosis and transmission index. This study was aimed to estimate CD4 cells and plasma HIV-1 RNA viral load among antiretroviral treatment (ART)-naive populations in three HIV treatment centres in Nigeria. Methods: We conducted a cross-sectional hospital-based study of 50 adult ART-naive patients. Whole blood and plasma samples were estimated for CD4 cells and HIV RNA-1 plasma viral load respectively. Results: The median age of the study participants was 35 years and 64% were female. The median CD4 cell count was 176 cell/μl while the median HIV viral load was 158391 copies/mL. There was a significant moderately strong, negative Spearman correlation between HIV-1 plasma viral load and CD4 cell count (r = −0.5007, P = 0.0002). Female recorded relatively higher CD4 cell count and lower plasma viral load. Six percent (6%) of the ART-naïve patients had undetectable viral load. Conclusion: This study indicates the baseline plasma viral load and CD4 cell count which can affect prognosis, disease progression and transmission. The drug-naïve participants reported with undetectable plasma RNA could be ‘elite’ controllers.

Keywords: ART-naïve, CD4 cell count, plasma viral load


How to cite this article:
Tahir MI, Aminu MA, Suleiman AB, Opaluwa AS, Ahmad AE, Anka AU. HIV-1 plasma RNA viral load and CD4 cell count in drug-naïve HIV-1 infected patients in Kaduna State, Nigeria. J Med Trop 2021;23:46-51

How to cite this URL:
Tahir MI, Aminu MA, Suleiman AB, Opaluwa AS, Ahmad AE, Anka AU. HIV-1 plasma RNA viral load and CD4 cell count in drug-naïve HIV-1 infected patients in Kaduna State, Nigeria. J Med Trop [serial online] 2021 [cited 2021 Sep 19];23:46-51. Available from: https://www.jmedtropics.org/text.asp?2021/23/1/46/314849




  Introduction Top


Human immunodeficiency virus type 1 (HIV-1) is the causative agent of acquired immunodeficiency syndrome (AIDS). It is characterized by extensive and dynamic genetic diversity, generating variants falling into distinct molecular subtypes as well as recombinant forms; these forms display an uneven global distribution. This diversity has implications for our understanding of viral transmission, pathogenesis, and diagnosis and profoundly influences strategies for vaccine development.[1]

Nigeria, the most populous country in Africa, was ranked second in the world for the highest HIV-1 burden.[2] It was estimated that 3.2 million people in Nigeria were living with HIV/AIDS, 220 000 new infections and 160 000 AIDS-related deaths in 2017.[3]

In Nigeria, initiation of antiretroviral therapy (ART) was conventionally based on baseline CD4 cell count and WHO clinical staging until recently where WHO treatment guideline became mainly ‘test and treat’ and patients tested positive are placed on Highly Active Antiretroviral Therapy (HAART) regardless of the CD4 cell count or WHO clinical staging.[4] Viral load and CD4 cell count are important markers for monitoring patients’ management.[5] These markers are necessary in estimating disease progression and prognosis.[6] Additionally, viral load present measure of infectivity and therefore, individuals with low viral loads will have low heterosexual transmission rate.[7] Our study reports distribution of CD4 cell count and HIV-1 RNA plasma viral load among ART-naïve HIV infected individuals in three testing centers in Kaduna State, Nigeria.


  Materials and methods Top


A total of 50 HIV infected drug-naïve adult participants were enrolled in this study from three voluntary counseling and testing (VCT) centers in Kaduna State, Nigeria. These participants were tested positive to HIV from April to October, 2018. The study was a cross-sectional hospital based and probability proportional to size (PPS)[28] sampling method was adopted [Table 1].
Table 1: Distribution of sample size across the three selected hospitals in Kaduna State, Nigeria

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Calculated sample size per study site total required sample size

Where a, b and c are the number of HIV infected individuals enrolled at various study sites in a six months period.

Basic demographic data of each volunteer was captured using structured questionnaire and transcribed to electronic questionnaire using Epi Info® version 7.2.2.2.6 software.

Nine milliliters (9ml) of venous blood was drawn from each participant into ethylene diamine tetraacetic acid (EDTA) anticoagulated vacutainer tube. One milliliter (1ml) of the well mixed whole blood was transferred to a 2ml-container for CD4 cells count. The remaining 8ml blood was centrifuged at 500Xg for 5 minutes and the separated plasma was transferred using clean Pasteur pipette into cryovials for plasma viral load estimation and other molecular testing.

CD4 cell enumeration

CD4 cell enumeration was carried out using BD FACSCount™ (BD Biosciences, USA) flow cytometry system.

Plasma HIV-1 Viral Load Estimation

Plasma HIV-1 RNA was quantified using COBAS® TaqMan® HIV-1 Test (Roche Molecular Systems, Inc., USA).

Ethical approval and individual informed consent

The study was conducted in accordance with the Declaration of Helsinki, and letter of ethical approval was obtained prior to commencement of study from the Health Research Ethics Committees of Ministry of Health, Kaduna State and Barau Dikko Teaching Hospital, Kaduna with reference number MOH/ADM/744/VOL.1/471 and 18-0001, respectively. Individual informed consent was sought from each volunteer prior to sample collection. Volunteers gave verbal and written consent after comprehending what the research was all about.


  Results Top


Results of CD4 cell enumeration among drug-naïve HIV infected participants in Kaduna state, Nigeria

The median CD4 cells among the participants was found to be 176.0 cell/μl (IQR: 74.75–402.8) with the minimum CD4 cell count of 8 cell/μl and the maximum of 900 cell/μl. Female had median CD4 cell count of 191.50 (IQR: 58.50-480.50) cell/μl whereas male had median CD4 cell count of 155 (IQR: 83–286) [Figure 1] and [Figure 2]. This apparent difference between the two means was not statistically significant (P = 0.7850) by Mann-Whitney test. The results of CD4 cell count alongside the age groups were presented in [Table 2]. There was no significant different in CD4 cells count across the age groups (P = 0.9371). Most of the participants, 17 (34%) had a CD4 cell count that falls between 51 and 200 cell/μl. Nine (18%) had CD4 cell count of ≤50 cell/μl while 7 (14) had CD4 cell count > 500 cell/μl [Table 3].
Figure 1: Box and whisker plot depicting CD4 cell count by gender

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Figure 2: Distribution of Viral Load inrelation to gender

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Table 2: Result of the CD4 cell count in relation to age groups of drug-naïve HIV infected participants in Kaduna State

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Table 3: Distribution of CD4 cell count of drug-naïve HIV infected participants in Kaduna State

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Results of HIV-1 plasma viral load among drug-naïve HIV-1 infected participants in Kaduna state, Nigeria.

Of the 50 drug-naïve HIV-1 infected participants, 47(94%) had detectable plasma levels of HIV-1 RNA. The median (IQR) HIV-1 plasma viral load among the participants were 158391 (44730- 548281) copies/ml and 651396 (1517000) copies/ml respectively. Female had median (IQR) and mean (SD) HIV-1 plasma viral load of 134619 (29324–700757) and 642331(1415000) respectively while their male counterparts had median (IQR) of 222221 (44730–452113) and mean (SD) HIV-1 plasma viral load 667512 (1727000) respectively. There was no significant difference between female and male viral load (P > 0.999). There was no significant different in HIV-1 plasma viral load across the age groups (P = 0.5203) [Table 4]. Five (10%) participants had plasma viral of ≤1000 copies/ml and 30 (60%) of the participants had plasma viral load of >100000 copies/ml [Table 5].
Table 4: Result of the HIV-1 plasma viral load in relation to age groups of drug-naïve HIV infected participants in Kaduna State

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Table 5: Distribution of plasma viral load of drug-naïve HIV infected participants in Kaduna State

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Correlation between CD4 cell count and HIV-1 RNA plasma viral load

There was a significant moderately strong, negative correlation between HIV-1 plasma viral load and CD4 cell count (Spearman correlation coefficient = −0.5007, P = 0.0002) [Figure 3]. [Table 6] presents the summary of cell count and HIV-1 RNA plasma viral load.
Figure 3: Correlation of CD4 cell count and HIV-1 plasma Viral Load

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Table 6: Data summary of CD4 cell count and plasma HIV-1 viral load among drug-naïve HIV-1 infected participants in Kaduna state, Nigeria

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  Discussion Top


The median CD4 cell count for the drug-naïve HIV-1 infected participants in this study was 176.0 cells/μl. This low median CD4 cell count is expected as the study participants were drug-naïve. Decreased CD4 count was reported to increase risk of clinical events and developing resistance.[8] Opportunistic infections may set in among the participants due to this low median CD4 cell count. In 2013, Anude et al.[9] in Nigeria reported comparable median baseline CD4 cell count of 142 cells/ μl. Lawn et al.[10] have reported a relatively lower baseline median CD4 cell count of 97cells/ μl. Govender et al.[11] reported in Soweto of South Africa a median CD4 cell count of 364 cells/μl. This finding demonstrated that female had higher median CD4 cell count than male; a result in agreement with several reports[5],[12],[11],[13] although our comparison had no statistical significance difference. The gender variation of the CD4 cell count has been linked to hormonal differences such as estradiol.[14].

Most of the participants, 26 (52%) had CD4 cell count < 200 cells/ μl and ART initiation at lower CD4 cell count has been associated with early mortality primarily from Immune Reconstitution Inflammatory Syndrome (IRIS).[15] Akinsegun et al.[5] reported similar result of 51.9% among ART-naïve HIV patients.

A total of 47 (94%) study participants had detectable levels of plasma HIV-1 RNA. The high percentage is not unconnected to the study population category as drug-naïve HIV infected individuals mostly have detectable plasma HIV level.[16] Previous studies have reported relatively lower proportion of study participants with detectable levels of plasma HIV-1 of 90.5%[16] in Nigeria and 91.5%[17] in India. The participants with undetected plasma RNA could be HIV or ‘elite’ controllers which are defined as drug-naïve HIV-infected individuals having undetectable viremia by standard assays (<50 copies RNA/mL).[18],[19] These patients with undetectable viral load could have been affected by social desirability bias and concealed their real ART status. The success rate for drug resistance testing decreases with low-level viremia as polymerase chain reaction fails to amplify the region of interest for sequencing reaction.[20],[21] Plasma with undetectable HIV RNA copies could pose challenge of unsuccessful nucleic acid amplification of target region. Undetectable viremia or plasma HIV-1 RNA levels below 1500 copies per milliliter were not associated with heterosexual HIV transmission, whereas the risk of transmission increased substantially with increasing viral loads.[22]

We reported in this study a median viral load of 158,391 copies/ml. Odaibo et al.[16] had reported higher mean of 450,370 copies/ml in Nigeria. Three different South African and a Botswana studies reported median plasma viral load of less than 100,000 copies/ml among drug-naïve patients.[23],[11],[6],[24] This notable difference could be linked to the various assays used for the plasma RNA quantification. High baseline plasma viral load can affect prognosis, disease progression and transmission.[22],[24]

This study reported female to have lower median HIV-1 plasma viral load than their male counterparts. This reciprocates the CD4 cell count where female had higher value than male. This data suggest that women may be a low-risk group for HIV transmission.

Majority of the patients (84%) had viral load > 10,000 copies/ml and 60% had viral load > 100,000 copies/ml. In a study, slow rate of achieving viral suppression during treatment has been connected with patients with viral load of > 100,000 copies/ml [25]. Another study by Farahani et al.[24] reported that the hazard of developing composite outcome for the individuals with baseline HIV-1 RNA >10,000 copies/ml to be 2.3 times higher than that for those with baseline HIV-1 RNA <10,000 copies/ml. Higher viral load in patients was found to deplete CD4 cells much faster than in those with relatively lower viral load.[24] In similar drug-naïve studies by Rangarajan et al.[26] and Farahani et al.[24] 60% and 53% of the study participants had viral load of > 10,000 copies/ml respectively. Additionally, Rangarajan et al. [26] reported 16% patients that had viral load > 100,000 copies/ml.

There was a strong negative correlation between HIV-1 plasma viral load and CD4 cell count in this study. This reciprocal association could be for the reason that plasma viral load strongly predicts the rate of decrease in CD4 cell count and progression to AIDS although, correct prognosis of HIV infected individual is better known when both CD4 cell and plasma viral load are used.[27] Rangarajan et al.[26] and Govender et al.[11] reported similar relationship however, Farahani et al.[24] reported negative but weak correlation.

This was a cross-sectional hospital-based study where 50 adult ART-naïve HIV infected persons were recruited by probability proportional to size (PPS) sampling method from three selected hospitals in Kaduna State, Nigeria. Samples were collected for CD4 cell count and viral load from the eligible participants and assayed using flow cytometry and real time PCR techniques. Data were collected using Epi-info® 7 and analysed using GraphPad Prims 6 statistical software package.


  Conclusion Top


Determining the CD4 cell count and plasma HIV viral load has strong implication for disease prognosis and management. Further study is recommended on the HIV-1 infected drug-naïve individuals with undetectable viral load to ascertain viral set points and other host and viral immunological indices.


  Limitation Top


Social desirability bias might have affected this study as patients could present themselves as drug-naïve while they must have been enrolled in another centre and loss to follow-up or relocated.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]



 

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