Browsing by Author "Mkhize-Kwitshana, Zilungile Lynette."

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Cytokine immune response profiles during 5 intestinal helminths and Mycobacterium 6 tuberculosis coinfection: An in vitro and human ex vivo study in KwaZulu-Natal.
(2023) Bhengu, Khethiwe Nomcebo.; Mkhize-Kwitshana, Zilungile Lynette.; Singh, Ravesh.; Naidoo, Pragalathan.
Background: There is a striking geographic overlap between helminths and tuberculosis (TB), particularly in developing countries like Africa. Underprivileged communities are more susceptible to these illnesses due to poverty, poor sanitation, and other environmental factor Helminth and tuberculosis infections exhibit distinct immune responses, which may be antagonistic in coinfected hosts and lead to poor prognosis. Helminth infections induce anti422 inflammatory Th2/Treg responses contrary to the pro-inflammatory Th1 responses triggered by Mycobacterium tuberculosis (Mtb) infection. Reduced TB protection has been associated with a strong Th2 response. Uncertainty exists on how helminth infection affects the host’s resistance to TB. This necessitates further investigation of immune responses in helminths and TB coinfection cases, particularly in KwaZulu-Natal (KZN). Aim: To determine the cytokine response profiles during intestinal helminth and TB coinfection using lymphocytic Jurkat and monocytic THP-1 cell lines for the in vitro study and TB and helminth coinfected South African adults for the human ex vivo study. Methods: Lymphocytic Jurkat and monocytic THP-1 cell lines were stimulated for 24 and 48 hours with Mtb H37Rv and Ascaris lumbricoides (A. lumbricoides) excretory-secretory protein extracts for the in vitro study. A cross-sectional study on consenting adult participants (≥18 years) (n = 414) recruited from primary health care clinics was conducted between March 2020 and August 2021 in Durban, KwaZulu Natal, for the pilot human ex vivo study. Blood and stool samples were collected from the recruited participants. The Kato-Katz and Mini-Parasep faecal parasite concentration techniques were used to detect intestinal parasite infections in stool samples. Blood samples were analysed to determine A. lumbricoides-specific immunoglobulin E (IgE) and immunoglobulin G4 (IgG4) levels to improve microscopy sensitivity. In this study, cytokine analysis was undertaken for 164 participants; 96 were HIV infected and had to be excluded, leaving 68 eligible participants. The eligible individuals were subdivided into uninfected controls (no helminth and TB infection) (n = 18), helminth only infected (n = TB only infected (n = 6), and TB and helminth co-infected (n = 6) groups. Thereafter, for both the in vitro and ex vivo study, the gene expression profiles of the T helper type 1(Th1) and transcription factors [Interferon-γ (IFN-γ), Tumour necrosis factor-α (TNF-α), Interleukin-2 (ILxvii 2), Nuclear factor of activated T cells 2 (NFATC2), Eomesodermin 446 (eomes), T helper 2 (Th2) and transcription factors (Interleukin-4 (IL-4), Interleukin5 (IL-5, Transforming growth factor-β (TGF-β), T helper type 17 (Th17) (Interleukin-17 (IL-17), immune protein and proteases (Granzyme B, Perforin), Regulatory T cells (Tregs) (Interleukin-10 (IL-10) and Fork head box P3 (FoxP3)] and the uninfected controls, TB alone, helminth alone and coinfected groups were determined using RT-qPCR. Results: (i) In vitro study: TB-stimulated Jurkat cells had significantly higher levels of IFN-γ, TNF-α, Granzyme B, and perforin compared to unstimulated controls, LPS, A. lumbricoides, and A. lumbricoides plus TB costimulated cells (p<0.0001). IL-2, IL-17, Eomes, and NFATC2 levels were also higher in TB-stimulated Jurkat cells (p<0.0001). TB alone stimulated cells had lower IL-5 and IL-4 levels compared to A. lumbricoides alone stimulated and TB plus A. lumbricoides costimulated Jurkat and THP-1 cells (p<0.0001. A. lumbricoides alone stimulated cells had higher IL-4 levels compared to TB plus A. lumbricoides costimulated Jurkat and THP- 1 cells (p<0.0001). TGF-β levels were also lower in TB alone stimulated cells compared to TB plus A. lumbricoides costimulated cells. IL-10 levels were lower in TB stimulated Jurkat and THP-1 cells compared to TB plus A. lumbricoides costimulated cells (p<0.0001. (ii) Ex vivo study: Similar results were noted for both the in vitro and the ex vivo study, although the human study had a smaller sample size. Conclusion: Data suggest that helminths induce a predominant anti-inflammatory Th2 and Treg response which may downregulate critical proinflammatory Th1 responses crucial for TB protection.
Diversity of rodent-borne zoonotic pathogens at the human-animal-environment interface in Qatar.
(2021) Islam, Md. Mazharul.; Mkhize-Kwitshana, Zilungile Lynette.; Farag, Elmoubashar Abubaker Abd.
Rodents are the most diversified terrestrial mammals in the world. These animals assist with maintaining a healthy ecosystem through the soil structure modification, aeration, and hydration, although 5-10% are regarded as pests and carry zoonotic pathogens. Besides consumption and damage of our food and property, they are responsible for the transmission of several diseases, including plague, typhus, and leishmaniasis. Commensal rodents are the primary source of these pathogens because of their close proximity to humans. Qatar is a small country in the Arabian Peninsula. Four rodent species have been recorded in this country, that includes three commensal (Mus musculus, Rattus norvegicus, and Rattus rattus) and one wild (Jaculus jaculus) species. The zoonotic importance of rodents is yet to be explored. Knowing the pathogens originating from rodents is essential for early preparedness, prevention, and control. Therefore, the current study was undertaken on commensal rodents, rodentborne zoonotic pathogens, and the factors that are associated with pathogen prevalence among rodents, such as rodent sex, age, and trapping location in Qatar. A cross-sectional study was conducted between August 2019 and February 2020, which trapped rodents from different facilities, such as livestock and agricultural farms, bachelor and family accommodations, and industrial and commercial areas of Qatar. After studying the morphological and morphometric characters, blood samples, ectoparasites and visceral samples were collected from the captured rodents. Parasitic, bacterial, and viral pathogens were identified and characterized using gross, necropsy, microscopic, culture, biochemical, immunologic, and molecular methods. Descriptive statistics and univariate analysis were conducted to detect rodents, rodent-borne pathogens abundance, and the related risk factors. The study trapped 148 rodents, most of which were adults (n = 138, 93.2%, 95% CI: 87.92–96.71), and from livestock farms (n = 79, 49%, 95% CI: 41.02–57.65). R. norvregicus was the most prevalent (n = 120, 81%, 95% CI: 73.83–87.05), followed by R. rattus (n=24, 16%, 95% CI:10.68–23.16) and M. musculus (n=4, 3%, 95% CI: 0.74– 6.78) with an average body weight of 18.8 ± 2.2 gm, 264.3 ± 87.5 gm, and 130 ± 71.3 gm, respectively. This is the first morphologic and morphometric study of commensal rodents in Qatar and the Arabian Peninsula that detected the Qatari rodents are relatively smaller than those of Turkey, Tunisia, and Iran. About 63.5% of the rodents were infected with at least one of the 9 species of parasites, viz. Xenopsylla astia, Ornithonyssus bacoti, Hymenolepis diminuta, Taenia taeniaeformis, Capillaria annulosa, Strongyloides spp., Giardia spp., Toxoplasma gondii, Trypanosoma lewisi, and Leishmania spp. Helminths were the most prevalent (46.0%), followed by ectoparasites (31.8%) and protozoa (29.1%). Going by individual species prevalence, X. astia ranked the highest (31.8%), where the lowest prevalent parasite was C. annulosa (0.7%). The prevalence of H. diminuta was positively correlated (OR=4.13; p = 0.00) with the prevalence of X. astia. The study also identified thirteen bacterial species, namely Acinetobacter baumannii, Aeromonas salmonicida, Citrobacter freundii, Citrobacter koseri, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Hafnia alvei, Klebsiella pneumoniae, Providencia stuartii, Proteus mirabilis, Pseudomonas aeruginosa, and Salmonella enterica, from the intestine samples. The majority of the bacteria were E. coli (54.63%, 95% CI: 44.76-64.24), followed by P. mirabilis (17.59%, 95% CI: 10.94-26.10), and K. pneumoniae (8.33%, 95% CI: 3.88-15.23). The study detected 31.58% (6/19, 95% CI: 12.58-56.55) of the flea pools, and one (1/1) mite pool was positive with Rickettsia spp. S. enterica showed the highest antimicrobial resistance (100% resistant to 8 antimicrobials). The top resistant antimicrobials were from cephalosporin, followed by penicillin and tetracycline groups. E. coli (26.92%, 95% CI:11.57-47.97) and K. pneumonia (50%, 95% CI: 6.76- 93.24) were ESBL (extended-spectrum beta-lactamases) producers. The studied rodents are indicators of the presence and dispersal of zoonotic pathogens in Qatar. Urgent action is needed to prevent future spillover of these pathogens at the human-animal-environment interface. It is essential to understand the biology, epidemiology, and transmission dynamics of these pathogens. Farm biosecurity and integrated pest management approach should be implemented in the farm premises. Implementing the One Health approach to combat rodent-borne zoonoses in order to reduce the risk of the future epidemic in Qatar is strongly recommended.
Epidemiology and alternative approaches for SARS-CoV-2 testing within limited resources settings=Isifundo ngembangela nokusabalala kwezifo i-epidemiology nezindlela ezahlukile zokuhlolwa kwe-SARS-CoV-2 esimeni sokwesweleka kwezinsiza.
(2023) Duma, Zamathombeni.; Mkhize-Kwitshana, Zilungile Lynette.; Chuturgoon, Anil Amichund.; Ramsuran, Veron.
Background: In the context of the global battle to contain the rapidly mutating SARS-CoV-2, diagnostic testing for SARS-CoV-2 infection remains a challenge, particularly in low-middleincome countries (LMICs) due to low socioeconomic backgrounds. Concerningly, because less attention is paid to asymptomatic cases, particularly in LMICs with limited resources for SARSCoV- 2 testing, the virus is spreading silently in communities, and the majority of these individuals could be contributing to the resurgence of SARS-CoV-2 infection. This study aimed to determine the epidemiology and alternative approaches for SARS-CoV-2 testing within limited resources settings. Methods: A total sample size of 1335 residual patient samples from the Global Health Innovation (GHI) laboratory was used for the epidemiology study and methods comparison. Results and Discussion: Literature review showed that high income countries (HICs) test more frequently for SARS-CoV-2 infection, with a range of 113% to 146% higher than LMICs (1% to 43%). The present study demonstrated a higher proportion of asymptomatic cases (68%) among SARS-CoV-2 infected patients. Regarding the methods comparison for the detection of SARSCoV- 2, the evaluated alternative methods [three RNA extraction (Lucigen QuickExtract™ RNA Extraction Kit, Bosphore EX-Tract Dry Swab RNA Solution, Sonicator method and four commercial SARS-CoV-2 RT-PCR assay kits (Nucleic Acid COVID-19 Test Kit (SARS-CoV-2), abTESTM COVID-19 qPCR I Kit, PCL COVID19 Speedy RT-PCR Kit, and PCLMD nCoV One- Step RT-PCR Kit)] were found to be cheaper and faster. Conclusion: Notably LMICs are undertesting for SARS-CoV-2 infection compared to HICs, and there was a higher proportion of asymptomatic cases among SARS-CoV-2 infected patients in South Africa. This study suggests that using the above-mentioned cost-effective, quick, and accurate evaluated alternative methods for mass SARS-CoV-2 testing in routine diagnostic laboratories with limited resources can help to increase testing capacity for SARS-CoV-2 infection in LMICs. This means that the sooner SARSCoV- 2 infection control and prevention measures can be implemented to reduce community transmission. IQOQA Isendlalelo: Esimeni somzabalazo womhlaba jikelele wokuvimba i-SARS-CoV-2 eguquguquka ngokushesha, uvivinyo oluhlolayo lokutheleleka nge-SARS-CoV-2 luselokhu luyinselelo, ikakhulu emazweni anemiholo ephansi kuya kwephakathi nendawo (low-middle income countries - MICs) ngenxa yesizinda senhlalomnotho ephansi. Ngokuxakayo-ke, ngenxa yokunganakwa kokutheleleka okungenazimpawu, ikakhulu ama-LMIC anezinsiza zokuhlolela i-SARSCoV-2, igciwane liyanda buthule emiphakathini, futhi iningi lalaba bantu lingase libe nomthelela yokuvembuka kabusha kokuthelelana nge-SARSCoV-2. Lolu cwaningo lwaluhlose ukuthola imbangela nokusabalala kwezifo i-epidemiology nezinye izindlela zokuhlola i-SARSCoV-2 esimeni sokwesweleka kwezinsiza. Izindlela: Kwasetshenziswa isampula eliphelele lamasampula eziguli ezisilele eziyi-1332 avela emalaborethri akwaGlobal Health Innovation (GHI) ngokocwaningo lwe-ephidemiyoloji nokuqhathaniswa kwezindlela. Imiphumela nengxoxo: Imibhalo eyabuyekezwa yakhombisa ukuthi amazwe anemiholo ephezulu (high income countries - HICs) ahlolela ukutheleleka kwe-SARS-CoV-2 kaningana ngokwebanga eliyi-113% kuya kweliyi-146% ngaphezu kwama-LMICs (1% kuya kuma-43%). Lolu cwaningo lwakhombisa isibalo esiphezulu lotho olungenazimpawu (68%) phakathi kweziguli ezitheleleke nge-SARS-CoV-2. Mayelana nezindlela zoqhathaniso lokuhlolwa kwe-SARS-CoV-2, izindlela ezahlukile ezihloliwe zokukhishwa kokuthathu (Lucigen QuickExtract™ RNA Extraction Kit, Bosphore EX-Tract Dry Swab RNA Solution, nendlela iSonicator) nezinsiza ze-assayi i-SARS-CoV-2 RT-PCR (Nucleic Acid COVID-19 Test Kit (SARS-CoV-2), abTESTM COVID-19 qPCR I Kit, PCL COVID19 Speedy RT-PCR Kit, ne-PCLMD nCoV OneStep RT-PCR Kit)] okwatholwa kushibhile futhi kushesha. Isiphetho: Ngokuqaphelekayo ama-LMIC akuhlolela kancane ukutheleleka nge-SARS-CoV-2 uma kuqhathaniswa nama-HIC, futhi kwakunesibalo esiphezulu sotho olungenazimpawu phakathi kweziguli ezitheleleke nge-SARS-CoV-2 eSouth Africa. Lolu cwaningo lukhomba ukuthi ukusebenzisa izindlela ezibalwe ngenhla ezishibhile, ezisheshayo, futhi ezinembayo nezihloliwe futhi ezingezinye izindlela zokuhlola isisindo se-SARS-CoV-2 engukuhlola okwejwayelekile emalabhorethri anezinsiza ezingeziningi kungasiza ekukhuliseni ukukwazi ukuhlola kokutheleleka nge-SARS-CoV-2 ema-LMIC. Lokhu kuchaza ukuthi uma kungashesha ukulawulwa kokutheleleka nge-SARS-CoV-2 nezinyathelo zokuvikeleka kungenziwa ukunciphisa ukuthelelana emphakathini.
The influence of helminths on immune responses to HIV.
(2009) Mkhize-Kwitshana, Zilungile Lynette.; Walzl, Gerhard.; Taylor, Myra.
In South Africa, co-infection with HIV and intestinal parasites is a major challenge in disadvantaged communities who live in densely populated under-serviced urban informal settlements. This pilot cross sectional study evaluates the immunological effects of co-infection with Ascaris lumbricoides and Trichuris trichura on the immune response to HIV. The work was a substudy of a prospective double blind, placebo-controlled investigation to test whether regular deworming changes the immune profile of HIV positive individuals with concurrent helminth infection. The substudy has a cross sectional design and presents pilot data that defines immune profiles of HIV-1 positive individuals with and without gastrointestinal helminth (Ascaris lumbricoides and Trichuris trichura) infection. The hypothesis was that concurrent helminth infection adversely affects immune responses against HIV. It was conducted in an area of high helminth endemnicity and limited infrastructural resources. Individuals with known HIV infection were recruited from an HIV Support Group and HIV negative individuals residing in the same area (for demographic matching) were used for comparison. The substudy was to provide pilot data for future larger scale and possible interventional studies. The current work is limited by the cross sectional design, moderate sample size and practical challenges. The profile of lymphocyte phenotypes, viral loads, eosinophils, activation markers, expression of the nuclear proliferation antigen-Ki67 and activation regulator antigen CTLA-4 were analysed using flow cytometry in HIV positive and negative subgroups with or without helminth infection. The type-1, type-2 and inflammatory cytokines were analysed using multiplex cytokine array technology. These were correlated with immune responses to HIV. Non parametric statistics were used to describe differences in the variables between the subgroups. A major finding of the study was the result of the supplementary use of the serological marker, Ascaris lumbricoides-specific IgE in addition to the presence (or absence) of helminth eggs in stools to classify intestinal helminth infection status. Two significant outcomes of this measure were the enhancement of diagnosis of current or recent helminth infection and, more importantly, the distinction of different phenotypes of individuals who displayed different immunological responses to co-infection with HIV and helminths. The different helminth infection phenotypes are defined by stool egg positivity (egg⁺) or negativity (egg⁻) with either high or low Ascaris-specific IgE (IgEhi or IgElo) respectively. The four subgroups, egg⁺IgEhi, egg⁺IgElo, egg⁻IgEhi and egg⁻IgElo showed different interactions with regards to immune response to HIV. It should be noted that no Trichuris specific IgE tests are commercially available but that there is significant antigenic cross-reactivity with Ascaris antigen. The presence of helminth stool eggs and high Ascaris IgE (egg⁺IgEhi) was associated with the following characteristics: reduction in numbers of all lymphocyte populations, frequent eosinophilia, highly activated immune profiles, antigen specific proliferative hyporesponsiveness, impaired type 1 cytokine responses in unstimulated and antigen stimulated cells and increased TNFα levels. In HIV infected individuals, the egg⁺IgEhi helminth infection status was associated with lower but not significant CD4⁺ counts and higher viral loads. A strong negative correlation was observed between viral loads, CD4⁺ and CD8⁺ cells in this subgroup. Subgroups with high IgE (egg⁺IgEhi and egg⁻IgEhi) had elevated Th2 markers with lower CD4⁺ counts and higher viral loads in the HIV⁺ group. The inverse correlation between viral load and CD4⁺ counts found in all the HIV⁺ participants was strongest in these two subgroups. Individuals with parasite eggs in stool and low Ascaris IgE (egg⁺/IgElo) presented a modified Th2 profile. This subgroup had high absolute numbers of all lymphocyte subsets in both HIV⁻ and HIV⁺ groups with higher CD4⁺ counts in the HIV⁻ and lower viral load in the HIV⁺ groups as well as higher interferon gamma, lower IL-4 and higher IL-10. In conclusion, the results suggest that helminth infections may be associated with deleterious effects on the immune responses to HIV in certain groups of susceptible individuals. The underlying reasons for the different stool egg/Ascaris IgE combinations in settings with high exposure to helminthes is currently not clear but genetic predisposition and environmental factors could play a role. Future studies of helminth- HIV co-infection have to ensure adequate definition of helminth infection status by the use of both stool examination and measurement of helminth-specific IgE as the infection phenotype is associated with differential effects on HIV associated immune responses. This may also apply to co-infection with other pathogens, including tuberculosis. The long-term effect of helminth co-infection in HIV positive people was not assessed in this study but requires further studies.
The interaction between nutrition, immunity and coinfections with human immunodeficiency virus and intestinal parasites in South African adults: investigating the use of prealbumin as a tool for nutritional assessment.
(2019) Mkhize, Brenda Thabisile.; Mkhize-Kwitshana, Zilungile Lynette.; Thobakgale, Christina Fanesa.
Highly prevalent HIV and helminth single infections continue to plague a significant proportion of the South African population. The geographic overlap of these infections lands to the expectation that high prevalence of co-infection with HIV and intestinal helminths exists, although this data for the South African adult population is lacking. Each of these single infections has an impact on the immune system, resulting in impaired responses due to the chronic activation. Also, both infections have an impact on the nutritional status, which may affect the potency of the immune responses, further compromising the immunity. A potent immune system requires adequate nutrition. Obesity, a form of malnutrition may mask micro- and macronutrient deficiency. Furthermore, obesity may result in low-grade inflammation, which may result is dysregulated responses. Therefore, malnutrition may start a cyclical process that may further predispose to infection, which in turn may result in malnutrition, where the cause-and-effect thread between malnutrition, infection and immune deficiency is indiscernible. Based on this, it was hypothesized that the HIV-intestinal helminth co-infection may have a deleterious effect on the nutrition and immunity of affected individuals, which may accelerate HIV progression. Thus, the aim of the study was to investigate the interaction between HIV and intestinal helminth single and co-infection with nutrition and immunity in an adult population (n = 263) in KwaZulu-Natal, a province with high prevalence of both HIV and intestinal helminths infections. The study expected to find an association between the co-infection with lower micro- and macro-nutrient levels, higher HIV viral load, increased immune activation, increased gene expression of Th2 and Treg cytokine responses and decreased Th1 cytokine responses compared to those singly infected and those uninfected with HIV and intestinal helminths. However, the study found no significant association between HIV and intestinal helminth single or co-infection with micro- and macronutrient deficiency, although a general pattern of low intake of the nutrients was noted among the investigated cohort, who had a substantial proportion being overweight and obese. Difficulty in the assessment of nutritional status in the milieu of HIV and intestinal helminth co-infection, obesity and inflammation was noted. Furthermore, HIV-intestinal helminth co-infection was associated with an antiviral cytokine response profile of highly expressed IFN-γ and TNF-α cytokine genes and reduced viral load. The co-infected individuals with the IgEhiIgG4hi intestinal helminth infection phenotype had a compromised immune profile of low CD4 counts. We recommend that antihelminthic interventions are included in the HIV management programmes, particularly in adults.