Browsing by Author "Singh, Shoohana."

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Cell signalling of the epidermal growth factor (EGF)/epidermal growth factor receptor (EGFR) axis in HIV associated pre-eclampsia.
(2021) Laldeo, Arisha.; Naicker, Thajasvarie.; Singh, Shoohana.
Background: Epidermal growth factor is a protein which, when bound to epidermal growth factor receptor, facilitates cell proliferation and differentiation, hence is vital for a successful normal pregnancy. In preeclampsia (PE), EGFR signalling is dysregulated leading to decline in blood flow to the fetus. Furthermore, aberrant EGF/EGFR signalling leads to deficient trophoblast development. The Trans-Activator of transcription (Tat) protein, displayed in HIV inhibits EGF related processes. Since PE and HIV infection are the leading causes of both maternal morbidity and mortality in South Africa; this study focuses on examining EGF/EGFR signalling in HIV associated PE and their effect on downstream targets. Methods: Post ethics approval; this study selected 80 pregnant women from an archive of retrospectively stored serum samples. The samples were stratified by type of pregnancy and HIV status into the following groups: a) HIV negative preeclamptic women (n=20), b) HIV positive preeclamptic women (n=20), c) HIV negative normotensive pregnant women (n=20) and d) HIV positive normotensive pregnant women (n=20). Both EGF and EGFR were multiplexed in a Bioplex immunoassay technique to determine their serum concentration across study groups at term. Results: Based on the clinical data, statistically significant differences were obtained across groups for gestational age (p < 0.001), birth weight (p < 0.001), systolic BP (p < 0.001), BMI (p = 0.048), diastolic BP (p < 0.001) and maternal weight (p = 0.002). However, no statistical significance was observed for maternal age across all study groups (p = 0.065). A significant decline in EGF levels were observed in PE compared to normotensive pregnancy, regardless of HIV status (p = 0.0214). Based on HIV status, no statistical significance in EGF concentration was observed (p = 0.6593). EGFR was significantly elevated in normotensive pregnant compared to preeclamptic women (p < 0.0001) (Figure 2A). In contrast, there were no significant differences of EGFR based on HIV status alone (p = 0.2092) (Figure 2B). However, a significant up-regulation of EGFR was observed between normotensive HIV positive compared to PE HIV positive women. Normotensive HIV negative was also significantly up-regulated compared to PE HIV positive (p < 0.0001). Normotensive HIV positive was significantly higher than PE HIV negative. Also, of note within the PE group, HIV negative EGFR levels were significantly up-regulated compared to the HIV positive group. Conclusion: This novel study outlines a significant down-regulation of EGF and EGFR in PE compared to normotensive pregnant women; regardless of HIV status. No significant differences were observed based on HIV status alone for serum EGF levels. This could be due to immune reconstruction as all HIV infected patients received HAART, hence may have neutralised EGF levels. Furthermore, these findings may be attributed to the Trans-Activator of transcription (Tat) protein which prevents EGF related function. However, for serum EGFR concentration there were significant differences within PE group based on HIV status. Significant differences were observed between normotensive and preeclamptic based on HIV status, except for normotensive HIV negative vs PE HIV negative. Furthermore, there was no significance in the normotensive group based on HIV status. The decreased levels of serum EGF/ EGFR could possibly be used as a biomarker for PE development during pregnancy. Isendlalelo: I-Epidermal growth factor iyiphrotheni okuthi uma ihlangene ne-epidermal growth factor receptor, isiza ngokwandisa amaseli nokuwehlukanisa, yingakho ibalulekile ekukhulelweni okujwayelekile okuphumelelayo. Ku-preeclampsia (PE), ukusayinda kwe-EGFR kuyaphazamiseka okuholela ekwehleni kokugeleza kwegazi ku-fetus. Ngaphezu kwalokho, ukusayina kwe-EGF / EGFR ngendlela eyehlukile kunalena eyejwayelekile kuholela ekushodeni kwe-trophoblast. I-Trans-Activator of transcription (Tat) protein ivimbela izinqubo ezihlobene ne-EGF uma ihlangene neHIV. Njengoba izifo ze-PE kanye ne-HIV kuyizimbangela ezihamba phambili zokugula nokufa komama eNingizimu Afrika; lolu cwaningo lugxile ekuhloleni ukusayina kwe-EGF / EGF-R ku-PE ehlobene ne-HIV kanye nomphumela wazo uma zixhuma endaweni eziyihlosile maphansi neseli. Izindlela: Ngemva kokugunyazwa kokuziphatha; lolu cwaningo lukhethe abesifazane abakhulelwe abangama-80 kungobo yomlando yamasampula e-serum agcinwe ngokudlule. Amasampula ahlukaniswa ngohlobo lokukhulelwa kanye nesimo se-HIV emaqenjini alandelayo: a) abesifazane abakhulelwe abangenayo i-HIV kodwa abanomfutho wegazi ophakeme (n=20), b) abesifazane abakhulelwe abane-HIV nomfutho wegazi ophakeme (n=20), c) abesifazane abakhulelwe abangenayo i-HIV futhi abanomfutho wegazi ojwayelekile (n=20), kanye d) nabesifazane abakhulelwe abane-HIV kodwa banomfutho wegazi ojwayelekile (n=20). Kokubili i-EGF ne-EGF-R zacwaningwa ngendlela ye-Bioplex immunoassay ukuze kutholwe inani lweserum yazo kuwo wonke amaqembu ocwaningo ngesikhathi. Imiphumela: Ngokubuka idatha esungulwe yimtholampilo, umehluko obalulekile ngokwezibalo watholwa kuwo wonke amaqembu eminyaka yobudala (p <0.001), isisindo sokuzalwa (p <0.001), i-systolic BP (p <0.001), BMI (p = 0.048), i-diastolic BP (p <0.001) kanye nesisindo sikamama (p = 0.002). Kodwa-ke awukho umehluko omkhulu ngokwezibalo owabonwa ngeminyaka yobudala bomama kuwo wonke amaqembu ocwaningo (p = 0.065). Ukwehla ngokwezibalo okubalulekile kwe-EGF kwabonwa i-PE uma kuqhathaniswa nokukhulelwa kwe-normotensive, kungakhathaliseki isimo se-HIV (p = 0.0214). Ngokubuka isimo se-HIV, akukho ukubaluleka kwezibalo kwe-EGF okuphawulwe (p = 0.6593). Kungakhathalekile ukuthi ngabe i-HIV ithini, i-EGFR iphakanyiswe kakhulu kumanormotensive abakhulelwe ngokuqhathaniswa nabesifazanebe-preeclamptic (p < 0.0001). Ngokuphambene, kwakungekho umehluko ophawulekayo we-EGFR ngokusekelwe esimweni se-HIV (p = 0.2092). Kodwa-ke, ukulawulwa okubalulekile kwe-EGFR kwabonwa phakathi kwe-HIV positive normotensive uma kuqhathaniswa nabesifazane abakhulelwe abangenayo ixvi HIV (p = 0.001); ngaleyo nkathi ukwehla okubalulekile kwe-EGFR kwaboniswe kuwo wonke amaqembu ocwaningo (p = 0.001). Isiphetho: Lolu cwaningo lwenoveli luveza ukwehla okubalulekile kwe-EGF ne-EGFR ku-PE uma kuqhathaniswa nabesifazane abakhulelwe abajwayelekile; kungakhathaliseki isimo se-HIV. Awukho mehluko omkhulu obonwe ngokusekelwe esimweni se-HIV. Lokhu kungase kube ngenxa yokwakhiwa kabusha kwamasosha omzimba njengoba zonke iziguli ezine-HIV zithole i-HAART, yingakho kungase kulinganise amazinga e-EGF/EGFR. Ngaphezu kwalokho, lokhu okutholakele kungase kubalulwe ku-Trans-Activator of transcription (Tat) protein evimbela umsebenzi ohlobene ne-EGF. I-down-regulation ye-EGF/EGFR ingase isetshenziswe njengokuhlola inkomba yokubikezela ukuthuthukiswa kwe-PE ngesikhathi sokukhulelwa.
Placenta progesterone and its receptor in HIV-associated pre-eclampsia.
(2022) Sewnarain, Serisha Azaria.; Naicker, Thajasvarie.; Singh, Shoohana.
Background: The maintenance of a healthy pregnancy is dependent upon the placental production of progesterone, which interacts with progesterone receptors (PR) to stimulate trophoblast invasion. Pre-eclampsia (PE) is associated with defective trophoblast invasion, and due to the high prevalence of HIV infection and pre-eclampsia in South Africa, this study examined the expression of placental progesterone and PR in HIV-infected women with PE. Methods: Placental tissue from 180 women were grouped into normotensive (N) (n = 60) and PE (n =120). The PE group was further stratified by gestational age into early-onset pre-eclampsia (EOPE) and late-onset pre-eclampsia (LOPE) (n = 60 per group). Both normotensive and PE groups were stratified by HIV status (HIV positive+ and HIV negative-) into N- (n=30), N+ (n=30), EOPE- (n=30), EOPE+ (n=30), LOPE- (n=30) and LOPE+ (n=30). Immunohistochemistry and morphometric image analysis were used to assess placental progesterone and PR immuno-expression in exchange and conducting villi. The Mann Whitney test was used to compare the effects of pregnancy type (normotensive vs. pre-eclamptic), HIV status (HIV+ vs. HIV-) and PE subtype (EOPE vs LOPE). For comparative analysis across all six study groups, a one-way ANOVA non-parametric Kruskal-Wallis test was used, followed by Dunn's Multiple Comparisons test. A two-way ANOVA was used to compare villi type (exchange vs conducting) and pregnancy type. Results: Progesterone was immunoexpressed within endothelial, mesenchymal and trophoblast cells within conducting and exchange villi whilst PR was mainly expressed on cytotrophoblasts and syncytiotrophoblasts. Progesterone and PR immuno-expression in exchange villi were significantly lower in the following groups: PE compared to the normotensive group (p = <0.0001 and p = <0.0001, respectively) and EOPE compared to the LOPE group (p = <0.0001 and p = <0.0001). Progesterone immuno-expression in the HIV+ group compared to the HIV- group was significantly lower (p = <0.0001), whilst PR expression was non-significant (p = 0.4291). Progesterone and PR immuno-expression in conducting villi were downregulated in the following groups: EOPE group compared to the LOPE group (p = <0.0001 and p = <0.0001) and in the HIV+ group compared to the HIV- group (p = <0.0001 and p = 0.0009). Progesterone immunoexpression was higher in the PE group compared to normotensive (p = 0.0326) and PR immunoexpression was non-significant (p = 0.6935). xiii There was a significant difference in progesterone and PR in exchange vs conducting villi (p = <0.0001 and p = <0.0001, respectively) and villi type accounted for 34.47% and 15.28% of total variance for progesterone and PR, respectively. Conclusion: This study observed a reduction in progesterone and PR immunoexpression in the exchange villi of pre-eclamptic placenta. Progesterone and PR immuno-expression were also significantly reduced in HIV+ placentas, with the EOPE+ group displaying the lowest immunoexpression. We postulate that HIV infection combined with cART may cause mitochondrial dysfunction that compromises progesterone synthesis. Progesterone deficiency results in minimal binding to PRs, which affects signalling pathways (PI3K/AKT, JAK-STAT, and MAPK cascades) and impairs trophoblast invasion. Notably, the EOPE group has the lowest immunoexpression of progesterone and PRs which links the downregulation of progesterone to defective placentation. This study links HIV infection to reduced progesterone production during pregnancy and associates decreased progesterone and PR immuno-expression with PE. Isendlalelo: Ukugcinwa kokukhulelwa okunempilo kuncike ekukhiqizweni kwe-placenta yeprogesterone, esebenzisana nama-progesterone receptors (PR) ukuze kuvuse ukuhlasela kwetrophoblast. I-Pre-eclampsia (PE) ihlotshaniswa nokuhlasela kwe-trophoblast enesici, futhi ngenxa yokusabalala okuphezulu kokutheleleka nge-HIV kanye ne-pre-eclampsia eNingizimu Afrika, lolu cwaningo luhlole ukuvezwa kwe-placental progesterone kanye ne-PR kwabesifazane abane-HIV abane-PE. Izindlela: Izicubu ze-placental ezivela kwabesifazane abangu-180 zahlanganiswa zaba yinormotensive (n = 60) ne-PE (n = 120). Iqembu le-PE laphinde lahlukaniswa ngeminyaka yokukhulelwa yaba yi-PE yokuqala kanye ne-PE yokufika sekwephuzile (n = 60 iqembu ngalinye). Womabili amaqembu e-normotensive kanye ne-PE ahlukaniswa ngesimo se-HIV (HIV+ ne-HIV negative-) aba yi-N- (n=30), N+ (n=30), EOPE- (n=30), EOPE+ (n=30), LOPE - (n=30) kanye ne-LOPE+ (n=30). I-Immunohistochemistry kanye nokuhlaziywa kwesithombe semorphometric kwasetshenziselwa ukuhlola i-placenta progesterone kanye ne-PR immunoexpression ekushintsheni nasekuqhubeni i-villi. Imiphumela: I-progesterone yayingabonakali ngaphakathi kwamaseli e-endothelial, mesenchymal kanye ne-trophoblast ngaphakathi kokuqhuba nokushintshanisa i-villi ngenkathi i- PR iboniswa ikakhulukazi kuma-cytotrophoblasts nama-syncytiotrophoblasts. I-progesterone ne- PR immuno-expression ekushintsheni i-villi yayiphansi kakhulu kumaqembu alandelayo: I-PE uma iqhathaniswa neqembu le-normotensive (p = <0.0001 kanye ne-p = <0.0001, ngokulandelana) kanye ne-EOPE uma kuqhathaniswa neqembu le-LOPE (p = <0.0001 kanye p = <0.0001). I-progesterone immuno-expression eqenjini le-HIV+ uma iqhathaniswa neqembu le- HIV yayiphansi kakhulu (p = <0.0001), kuyilapho inkulumo ye-PR yayingabalulekile (p = 0.4291). I-progesterone ne-PR immuno-expression ekuqhubeni i-villi yehlisiwe emaqenjini alandelayo: Iqembu le-EOPE uma liqhathaniswa neqembu le-LOPE (p = <0.0001 kanye ne-p = <0.0001) naseqenjini le-HIV+ uma liqhathaniswa neqembu le-HIV (p = <0.0001 futhi p = 0.0009). Iprogesterone immuno-expression yayiphezulu eqenjini le-PE uma kuqhathaniswa nenormotensive (p = 0.0326) kanye ne-PR immuno-expression yayingabalulekile (p = 0.6935). xv Kunomehluko omkhulu ku-progesterone ne-PR ekuhwebeni ngokuqhudelana nokuqhuba i-villi (p = <0.0001 kanye ne-p = <0.0001, ngokulandelana) kanye nohlobo lwe-villi lubalelwa ku- 34.47% no-15.28% wokuhluka okuphelele kwe-progesterone ne-PR, ngokulandelana. Isiphetho: Lolu cwaningo lubone ukuncipha kwe-progesterone kanye ne-PR immunoexpression ku-villi yokushintshanisa ye-pre-eclamptic placenta. I-progesterone ne-PR immuno-expression nazo zehliswa kakhulu kuma-placenta e-HIV+, neqembu le-EOPE+ libonisa ukubonakaliswa okuphansi kokuzivikela komzimba. Sibeka umbono wokuthi ukutheleleka nge-HIV kuhlanganiswe ne-cART kungase kubangele ukungasebenzi kahle kwe-mitochondrial okuphazamisa ukwakheka kwe-progesterone. Ukuntuleka kwe-progesterone kubangela ukubophezela okuncane kuma-PRs, okuthinta izindlela zokubonisa (PI3K/AKT, JAK-STAT, kanye ne-MAPK cascades) futhi kulimaze ukuhlasela kwe-trophoblast. Ngokuphawulekayo, iqembu le-EOPE line-immuno-expression ephansi kakhulu ye-progesterone kanye ne-PRs exhumanisa ukulawulwa kwe-progesterone nokuzala okungalungile. Lolu cwaningo luxhumanisa ukutheleleka nge-HIV nokuncipha kokukhiqizwa kwe-progesterone ngesikhathi sokukhulelwa kanye nokuhlotshaniswa nokuncipha kwe-progesterone kanye ne-PR immuno-expression ne-PE.
Seroprevalence and viral quantification of Kaposi Sarcoma-associated Herpes Virus (KSHV) in a Human Immunodeficiency Virus (HIV) infected adult South African cohort.
(2017) Singh, Shoohana.; Mosam, Anisa.; Shaik, Fahmida.; Uldrick, Thomas S.; Naidoo, Kogieleum.
Background Kaposi sarcoma-associated herpes virus (KSHV), also known as human herpes virus 8 (HHV8), is aetiologically implicated in Kaposi’s sarcoma (KS). Although HIV associated KS has increased in incidence and is a public health problem in South Africa, serological studies of KSHV have not been extensively documented in this population. This cross-sectional study investigates the seroprevalence and viral load of KSHV in an adult South African cohort. Method Cross-sectional data of 140 participants attending an urban research HIV counseling and testing (HCT) clinic site in Durban, KwaZulu-Natal, between July and October 2013 was analyzed. Detection of antibodies against latent (Orf73) and lytic (K8.1) KSHV antigens was performed on 70 HIV-seropositive and 70 HIV-seronegative participants. Subjects reactive to either antigen were considered KSHV seropositive and analyzed for salivary KSHV DNA, which was quantified using primers for the K6 gene region. Results The demographic characteristics of the two groups were similar, with 36% males (median age, 35yrs.) and 64% females (med. age, 34yrs.) in the HIV-positive group, and 31% males (med. age, 36.5yrs.) and 69% females (med. age, 36.5yrs.) in the HIV-negative group. Of 70 HIV-positive participants, 100% were black Africans, as was 97% of the HIV-negative group, with the remaining 3% being Indian/Asian and Mixed race. Only 24% of HIV-positive patients were on Anti-retro viral treatment. Fifty-four percent of all participants tested positive for KSHV, with 33% reactive to lytic K8.1, 37% to latent Orf73 and 21% to both. Of those HIV-positive, 50% were seropositive for K8.1 and 46% for Orf73. In those HIV-negative, 16% were seropositive for K8.1 and 29% for Orf73. The HIV-positive group demonstrated a significantly higher percentage KSHV seropositivity (70% vs. 37%, p=0.0001). Amongst the KSHV seropositive participants, KSHV DNA was detected in 41 % HIV-positive and 23% HIV-negative participants. Conclusion KSHV seroprevalence was high in South African adults attending an urban HCT clinic. HIV positive status was associated with a higher KSHV seropositivity and a greater KSHV salivary shedding. HIV positive individuals should be tested for KSHV infection and those found infected, be monitored aggressively for development of KS.