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 Table of Contents  
ORIGINAL ARTICLE
Year : 2019  |  Volume : 4  |  Issue : 1  |  Page : 16-20

Immunochemical evidence of Trypanosoma cruzi and Toxoplasma gondii parasitic infections in human immunodeficiency virus/acquired immunodeficiency syndrome subjects in relationship with the CD4+ count


1 Department of Medical Laboratory Science, Edo University, Iyamho, Nigeria
2 Department of Medical Laboratory Science, Achievers University, Owo, Ondo State, Nigeria

Date of Submission17-Nov-2018
Date of Acceptance15-Dec-2018
Date of Web Publication20-Feb-2019

Correspondence Address:
Prof. Mathew Folaranmi Olaniyan
Department of Medical Laboratory Science, Edo University, Iyamho
Nigeria
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijas.ijas_19_18

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  Abstract 


Study Background: Trypanosoma cruzi and Toxoplasma gondii parasitic infections in human immunodeficiency virus (HIV)-positive patients could accelerate the progression of HIV infection, which may result into immunosuppression due to immune responses involving cytokines and CD4 + cells.
Aim and Objective: This work was designed to determine the immunochemical evidence of T. cruzi and T. gondii parasitic infections in HIV/acquired immunodeficiency syndrome subjects in relationship with the CD4+ count.
Materials and Methods: Fifty HIV-positive subjects (25 females and 25 males) aged 14–57 years were recruited as test subjects while 100 HIV-seronegative age-matched individuals were recruited as control. T. cruzi and T. gondii parasitic infections were determined in the subjects by ELISA, while CD4+ cells enumeration was carried out in the subjects by cyflowmetry, and HIV test was carried out by immunochromatography and western blotting.
Results: The results obtained showed a significantly lower CD4+ cells in T. cruzi- and T. gondii-infected HIV-positive patients than the HIV-positive and HIV-negative subjects who were not infected with any of the parasites (<0.05). There was also a significantly lower CD4+ cells in T. gondii-infected HIV-positive patients than T. cruzi-infected HIV-positive patients (<0.05). The frequency of T. cruzi in HIV-positive patients was 18% (9) which was more prevalent in male patients (12%[6] vs. 6% [3]) while that of T. gondii-infected HIV-positive patients was 14% (7), was more prevalent in females than males (10%[5] vs. 4% [2]). T. cruzi parasitic infection was found to be more prevalent compared to T. gondii infection among HIV-positive patients (18%[9] vs. 14% [7]).
Conclusion: There parasitic infection caused a significant decrease in CD4+ cells in HIV subjects. There was also a gender difference in the parasitic infection of T. cruzi and T. gondii while the frequency of T. cruzi was 18% (9) and T. gondii was 14% (7).

Keywords: CD4+ cells, human immunodeficiency virus, Toxoplasma gondii, Trypanosoma cruzi


How to cite this article:
Olaniyan MF, Kilo OA. Immunochemical evidence of Trypanosoma cruzi and Toxoplasma gondii parasitic infections in human immunodeficiency virus/acquired immunodeficiency syndrome subjects in relationship with the CD4+ count. Imam J Appl Sci 2019;4:16-20

How to cite this URL:
Olaniyan MF, Kilo OA. Immunochemical evidence of Trypanosoma cruzi and Toxoplasma gondii parasitic infections in human immunodeficiency virus/acquired immunodeficiency syndrome subjects in relationship with the CD4+ count. Imam J Appl Sci [serial online] 2019 [cited 2019 Sep 19];4:16-20. Available from: http://www.e-ijas.org/text.asp?2019/4/1/16/252592




  Introduction Top


Trypanosoma cruzi and Toxoplasma gondii are protozoan hemoparasites that cause parasitic infections known as trypanosomiasis and toxoplasmosis, respectively. They could elicit immune response leading to the production of cytokines that stimulate CD4 and CD8 for cell-mediated innate immunity. Human immunodeficiency virus (HIV) is the virus that causes acquired immunodeficiency syndrome (AIDS). When a person becomes infected with HIV, the virus attacks and destroys the CD4+ bearing cells.[1],[2],[3],[4],[5] Massive depletion of CD4+ bearing cells could lead to immunosuppression/AIDS.[6] T. cruzi is of parasitic euglenoids. It is a protozoan and T. cruzi can bore tissue in another organism and feed on blood including lymph which could result into diseases such as trypanosomiasis in humans (Chagas disease in South America), dourine and surra in horses, and a  Brucellosis More Details-like disease in cattle.[7] The vector or transmitting agent of T. cruzi is hematophagous insect triatomine (also known as assassin bug, cone-nose bug, and kissing bug). The triatomine while in human for shelter bites and sucks blood for food. Human contract T. cruzi when in contact with triatomine feces deposited on the host's skin surface and then bites. T. cruzi penetrates through the feces when the affected human scratches the bite area.[8],[9],[10] The course of trypanosomiasis in the affected individual progresses with the development of the trypanosome into a trypomastigote in the blood and into an amastigote in tissues. Acute of trypanosomiasis is usually without symptom though can cause localized swelling at the site of entry. The chronic form may develop within 30–40 years of infection and affect organs such as heart, esophagus, colon, and peripheral nervous system. T. cruzi-infected individuals may die from heart failure.[8],[9],[10]

Toxoplasmosis is a parasitic disease caused by T. gondii possibly with no obvious symptoms in adults. Occasional symptom include a few weeks/months of mild flu-like illness which include muscle aches and tender lymph nodes, and in few cases, eye problems may develop. T. gondii infection in those with low immunity severe symptoms such as seizures and poor coordination may occur. It could infect pregnant woman to cause congenital toxoplasmosis which may affect the child. Toxoplasmosis is usually spread by eating poorly cooked food that contains cysts of T. gondii contaminated by the feces of an infected cat or from infected mother to their child during pregnancy. The sexual reproduction of T. gondii takes place in cat and can infect most types of warm-blooded animals, including humans.[11],[12],[13],[14],[15]

AIDS is caused by a virus called HIV. The HIV disease depletes the immune materials, making affected individual susceptible to infections and diseases. This susceptibility increases with the progress of the HIV disease/syndrome. HIV is a virus that destroys immune cells known as CD4+ cells, which are a subset of T-cells.[1],[2],[3],[4],[5]

HIV/AIDS affects immune system which can make affected individuals susceptible to infections. There is little information on immunochemical evidence of T. cruzi and T. gondii parasitic infections in HIV/AIDS subjects in relationship with the immune materials such as CD4+ count hence the need for this work. This work was designed to determine the immunochemical evidence of T. cruzi and T. gondii parasitic infection in HIV/AIDS subjects in relationship with the CD4+ count.


  Materials and Methods Top


Materials

Study area

Federal Medical Centre, Owo, is a public health care center located in Owo, a city in Ondo State, Southwestern Nigeria. It is a tertiary institution training physician, medical laboratory scientists, and nurses. Owo is a city in Ondo State of Nigeria. Between 1400 and 1600 AD, it was the capital of a Yoruba city-state. The local government has a population of 222,262, based on the 2006 population census.

Study population

The sample size of this study was determined using the statistical formula:

Z2 × (p) × (1 − p) ÷ C2 (Bill Godden, January 2004).

SS = Sample Size

Z = Z-value A (e.g., 1.96 for a 95% confidence level)

P = Percentage of population picking a choice, expressed as decimal B

C = Confidence interval, expressed as decimal (e. g., 0.04 = ±4% points) A Z-value (cumulative normal probability table) represents the probability that a sample will fall within a certain distribution.

CD C (2016) reported a prevalence of HIV infection as 3.1% in Nigeria.

The calculated sample size was 47 but 50 HIV-positive subjects (25 females and 25 males) aged 14–57 years were recruited as test subjects why 100 HIV-seronegative age matched individuals was recruited as control.

Blood specimen

Venous blood sample was obtained from each of the subjects into EDTA specimen bottles for the determination of anti-HIV, parasitic infection, and CD4 enumeration.

CD4 enumeration

This was carried out by cyflowmetery using Partec CyFlow Counter and the complete solution for CD4 and CD4% testing.

The CyFlow Counter is a fully equipped compact and robust desktop flow cytometer with green laser excitation and three optical parameters (SSC and two fluorescence channels). It performs both fluorescence analysis and true volumetric absolute cell counting without the need for reference beads or an hematology counter. It represents a reliable solution with variable sample throughput for local heath care centers as well as district and regional hospitals. The CyFlow® Counter is simple to install and does not need any additional setup time. With the CD4/CD4% easy count kit reagents, blood sample preparation and incubation occur outside the device, which increases the sample throughput. Results are presented within 3 min as diagrams and as counting results in cells/μl blood sample or percentage.

Human immunodeficiency virus test

This was determined by immunochromatography and western blot methods.

Human immunodeficiency virus screening

HIV screening was carried out on the test and control volunteers after pretest counseling by using the reagent kit of Abbot Laboratories Co. Ltd, Japan. The Abott Determine HIV-1/2 is an in vitro, visually read qualitative immunoassay for the detection of antibodies to HIV-1 and HIV-2 in the human serum plasma or whole blood. The test is intended as an aid to detect antibodies to HIV-1/HIV-2 from infected individuals.

Human immunodeficiency virus confirmatory test

The HIV confirmatory test was carried out on all the volunteers by Western blot assay, using reagent kit of Immunoetics, Inc., 27 Dryclock Avenue, Boston, USA (http//www.immunoetics.com).

Principle the qualicodeHIV1/2 kit is a qualitative immunoblot assay based on the western blot principle. The assay is performed on immunoblot membrane containing HIV-1 viral lysate protein (HLTVIII B stain) and a recombinant HIV-2 protein. To produce the membrane HIV-1 viral protein are fractionated according to molecular weight on a polyacrylamide slab gel in the presence of sodium deodecyl sulfate. The separated HIV-1 is then transferred through electrophoretic blotting from the gel to a nitrocellulose membrane two bands are directly sriped on the membrane 1).

Immunochemical evidence of Trypanosoma cruzi and Toxoplasma gondii in the blood samples

This was determined immunochemically by ELISA.

Method of statistical analysis

Data collected were subjected to statistical analysis to determine level of significance at probability of 95% confidence limit using SPSS18.0.

Ethical consideration

Permission/approval was obtained from the ethical and research committee of Federal Medical Centre, Owo, Nigeria, where the study was carried out. The consent of each subject was obtained through a consent form.


  Results Top


The results obtained showed a significantly lower CD4+ cells in T. cruzi- and T. gondii-infected HIV-positive patients than the HIV-positive and HIV-negative subjects who were not infected with any of the parasites [<0.05; [Table 1], [Table 2] and [Figure 1]].
Table 1: Mean and standard deviation of CD4+ count and frequency of Trypanosoma cruzi and Toxoplasma gondii in the subjects

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Table 2: Comparative analysis of mean and standard deviation of CD4+ count in the subjects

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Figure 1: CD4+ cell count obtained in the subjects

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There was also a significantly lower CD4+ cells in T. gondii-infected HIV-positive patients than T. cruzi-infected HIV-positive patients (<0.05) [Table 1], [Table 2] and [Figure 1].

The immunochemical of T. cruzi infection in HIV-positive patients was 18% (9) which was more prevalent in male patients (12%[6] vs. 6% [3]) while that of T. gondii-infected HIV-positive patients was 14% (7) and was more prevalent in females than males (10%[5] vs. 4% [2]) [Table 1] and [Figure 2].
Figure 2: Comparative description of frequency of Trypanosoma cruzi and Toxoplasma gondii in the subjects

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T. cruzi parasitic infection was found to be more prevalent compared to T. gondii infection among HIV-positive patients (18%[9] vs. 14% [7]) [Table 1] and [Figure 2].


  Discussion Top


The results obtained showed a significantly lower CD4+ cells in T. cruzi- and T. gondii-infected HIV-positive patients than the HIV-positive and HIV-negative subjects who were not infected with any of the parasites.

This could be attributed to previous reports that one of the clinical manifestations of T. cruzi and T. gondii infection in HIV-positive patients is immunosuppression due to decrease in CD4+ cells. Most coinfected patients acquire T. cruzi through the vector-borne route in childhood and acquire HIV later in life. The clinical manifestations of Chagas disease with HIV/AIDS are associated with the central nervous system (74.2% of cases), probably as a result of severe immunosuppression that gives rise to the primary clinical manifestation of meningoencephalitis. T. gondii infection stimulates the production of IL-2 and interferon-gamma (IFN-γ) by innate immune system. The two cytokines elicit a CD4+ and CD8+ T-cell mediated immune response. IL-12 is produced during T. gondii infection to activate natural killer cells which can destroy CD4+ bearing cells infected with HIV, leading to depletion in CD4+ bearing cells[6] as it was found in this study.

There was also a significantly lower CD4+ cells in T. gondii-infected HIV-positive patients than T. cruzi-infected HIV-positive patients. The explanation to this is that mechanisms by which HIV induces susceptibility to opportunistic infections such as toxoplasmosis are likely multiple.[16],[17],[18],[19],[20],[21] These include depletion of CD4 T-cells; alterations in the production of IL-2, IL-12, and IFN-γ; and impaired cytotoxic T-lymphocyte immunological function. Cells from HIV-infected patients manifest decrease in in vitro generation of IL-12 and IFN-γ and decrease in the expression of CD154 in response to T. gondii infection. These inadequacies may contribute greatly to the development of toxoplasmosis associated with HIV infection.[16],[17],[18],[19],[20],[21]

T. gondii infection in HIV-infected patients has been associated with low CD4 T-cell counts.[16],[17],[18],[19],[20],[21] Long-term T. cruzi infection in humans might exhaust long-lived memory T-cells.[22]

The immunochemical evidence of T. cruzi infection in HIV-positive patients was 18% (9) which was more prevalent in male patients (12%[6] vs. 6% [3]) while that of T. gondii-infected HIV-positive patients was14% (7) and was more prevalent in females than males (10%[5] vs. 4% [2]). The immunochemical evidence of T. cruzi parasitic infection was found to be more compared to T. gondii infection among HIV-positive patients (18%[9] vs. 14% [7]). The coinfection of T. cruzi and HIV/AIDS has been documented widely (Carolina et al., 2014). Between 10% and 40% of HIV-infected patients in the United Sates have antibodies against T. gondii[17],[18],[19] which could be associated with the immunochemical evidence/frequency obtained in this study. Dormant parasitic infection could be reactivated when coinfected with HIV. Kamal and El Sayed Khalifa[23] found that those infected with parasites are more likely to be infected by HIV which may deplete CD4. However, it is disputed whether or not the viral infection is more severe because of the parasites.


  Conclusion Top


The results obtained showed a significantly lower CD4+ cells in T. cruzi- and T. gondii-infected HIV-positive patients than the HIV-positive and HIV-negative subjects who were not infected with any of the parasites. There was also a significantly decrease CD4+ cells in T. gondii-infected HIV-positive patients compared to the results obtained in T. cruzi infected HIV-positive patients. The immunochemical evidence of T. cruzi in HIV-positive patients was 18% (9) which was more in male patients (12%[6] vs. 6% [3]) while that of T. gondii-infected HIV-positive patients was 14% (7) and was more in females than males (10%[5] vs. 4% [2]). T. cruzi parasitic infection was found to be more prevalent compared to T. gondii infection among HIV-positive patients (18%[9] vs. 14% [7]).

Recommendation

Determination of T. cruzi and T. gondii in HIV/AIDS subjects in addition to CD4+ count will provide useful guide in the management of HIV/AIDS.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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Torgerson PR, Mastroiacovo P. The global burden of congenital toxoplasmosis: A systematic review. Bull World Health Organ 2013;91:501-8.  Back to cited text no. 14
    
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Montoya JG, Remington JS. Toxoplasma gondii. In: Mandell GL, Bennett JE, Dolin R, editors. Principles and Practice of Infectious Diseases. Philadelphia: Churchill Livingstone; 2000. p. 2858-88.  Back to cited text no. 19
    
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Subauste CS, Wessendarp M, Portilllo JA, Andrade RM, Hinds LM, Gomez FJ, et al. Pathogen-specific induction of CD154 is impaired in CD4+ T cells from human immunodeficiency virus-infected patients. J Infect Dis 2004;189:61-70.  Back to cited text no. 21
    
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Albareda MC, Olivera GC, Laucella SA, Alvarez MG, Fernandez ER, Lococo B, et al. Chronic human infection with Trypanosoma cruzi drives CD4+ T cells to immune senescence. J Immunol 2009;183:4103-8.  Back to cited text no. 22
    
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    Figures

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    Tables

  [Table 1], [Table 2]



 

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