Toll-like receptor 4 polymorphism influence on

the clinical course of chronic hepatitis C in HIV-infected

patients The presenting author is

Tetiana Kyrychenko Authors: G. Dubynska, T.Koval

Institutes: HIGHER MEDICAL EDUCATIONAL INSTITUTION

OF UKRAINE "UKRAINIAN MEDICAL STOMATOLOGICAL ACADEMY"

Regional HIV/AIDS Prevention and Control Center, Poltava Ukraine

Relevance Patients co-infected with HIV/HCV demonstrate the

increasing risk in developing cirrhosis and hepatocellular carcinoma (Harting H., 2011).

Recent studies have shown that TLR4 Asp299Gly SNP impacts the development of liver cirrhosis and on the efficiency of antiviral therapy in HCV-infected persons (Lorean A-R, 2012; Ahmed A. Al-Qahtani, 2014; Peric M, 2015)

Taking into account disputable findings obtained by these researchers and lack of information about the influence of TLR-4 polymorphism on the clinical course of chronic hepatitis C in HIV-infected persons, the research I am going to present, seems to be very relevant.

Fig.1. Schematic view of the Toll-like Receptor 4 (TLR4)

signaling pathway. (B. Ferwerda at all., 2007)

Objectives to study the prevalence of TLR4

Asp299Gly SNP in the healthy persons, HIV-positive persons and HIV/HCV co-infected persons in Ukraine;

to reveal clinical manifestations of chronic hepatitis C depending on TLR4 genotype variants.

to study the efficiency of antiviral therapy of chronic hepatitis C in HIV/HCV co-infected persons depending on TLR4 genotype variants

Study Subjects. The study involved 62 antiretroviral treatment-naive HIV/HCV co-infected patients, 106 HIV – positive persons and 131 healthy subjects. Laboratory procedures. Routine CD-4 cell count was carried out by conventional flow cytometry of fresh blood samples taken from all the patients. Plasma HIV-RNA and HCV -RNA were routinely measured by the polymerase chain reaction (PCR) (Real time HIV-1, Abbout). Fibrosis stages were determined by Fibrotest (BioPredictive France) TLR4 genotyping was performed by real-time PCR using oligonucleotide primers (Tercik PCR system, DNA-technology, Moscow) Statistical processing was carried out by using the SPSS/PC+ statistical package (version 17; SPSS, Chicago, IL, USA). Allelic and genotypic prevalence was estimated by direct counting and Pearson’s chi-squared (𝜒𝜒2 ) tests. The results were represented in terms of odds ratio (OR), 95% confidence intervals (95% CI) and 𝑃𝑃 values. Logistic regression model was also used.

Methods

Characteristics Wild type (Asp/Asp, n=50)

Asp299Gly heterozygous (Asp/Gly, n=12)

Male 30 (60) 7 (58,3)

Mean Ageª (years) ± standard deviation

35,31±0,51 35,43±1,33

Nadir CD-4 cell countª, mean cell/mm³± standard deviation

359,52±16,2 363,6±30,3

HIV Clinical stage (%) A B C

3 (6,0)

22 (44,0) 25 (50,0)

1 (8,3)

6 (50,0) 5 (41,7)

1 HCV genotype (%)

39 (78)

9 (75)

Fibrosis stage (%) 0-2 3-4

15 (30) 35 (70)

3 (25) 9 (75)

Description of a cohort of 62 patients infected with HIV/HCV and studied in relation to SNP of TLR4

(No. (%) of patients)

Хронический гепатит С

RVR EVR SVR

4 week 12 week 48 week stop After 24 weeks Start 24 week

DVR

Monitoring of therapy response to PEG-IFN plus ribavirin

Twenty HIV/HCV co-infected patients (Asp/Asp n=20, Asp/Gly n=5) were treated with standard interferon and ribavirin therapy (Peg-IFN 2b 1,5 mg/kg weekly +Ribavirin – 1000-1200 mg/day, 48 weeks), and their viral load was evaluated at 4,12, 24, 48, 72 weeks since the beginning of therapy.

Genotypic distribution of TLR4 SNP between healthy control subjects, HIV-positive patients, and HIV/HCV co-infected patients

* HIV+ and HIV/HCV+ compared to healthy, p

Prevalence of Symptoms in persons with HIV/HCV co-infection in relation to SNP of the Toll-like receptor 4 gene

(No. (%) of patients) Variable

Wild type (Asp/Asp,

n=50)

Asp299Gly heterozygous

(Asp/Gly, n=12)

OR (95% CI)

𝜒𝜒2 𝑃𝑃 value

Physical tiredness

20 (40,0) 9 (75,0) 20,45 (2,32-179,65)

4,76 0,007

Mental tiredness 30 (60,0) 11 (91,6) 20,31 (2,11-196,03)

4,3 0,009

Poor concentration

19 (38,0) 7 (58,3) 1,61

Irritability 19 (38,0) 6 (50,0) 0,60 Sleep problems 15 (30) 6 (50) 1,72

Abdominal pain 11 (22,0) 7 (58,3) 20,56 (2,01-210,03)

6,20 0,011

Nausea 14 (28) 6 (50) 2,14 Poor appetite 13 (26) 4 (33,3) 0,26 AST and ALT >N 21 (52,0) 11 (91,6%) 16,1

(1,27-201,3) 6,3 0,03

CD-4 cell/mm³

Asp299Gly TLR-4 The duration of HCV infection

Alcohol consumption

Liver fibrosis stage III-IV

τ = 0,36 τ = -0,47

τ= 0,37 τ = 0,34

The main factors influencing on the development of liver fibrosis

in HIV-infected patients

The development of hepatic fibrosis depending on TLR4 genotype

variants Error Bars show Mean +/- 1,0 SE

Dot/Lines show Means

2,00 4,00 6,00 8,00

Рік

2,00

3,00

4,00

Стадія

Error Bars show Mean +/- 1,0 SE

Dot/Lines show Means

2,50 5,00 7,50 10,00 12,50

Рік

2,00

2,50

3,00

3,50

4,00

Стадія

Years Years

Fibr

osis

sta

ge

Fibr

osis

sta

ge

Asp/Gly Asp/Asp

R = 0,51, p

SVR of Chronic Hepatitis C therapy in HIV/HCV co-infected persons

* Asp/Gly compared to Asp/Asp, p

Conclusion This study suggests greater risk for developing

HCV clinical features as physical and mental tiredness, abdominal pain and cytolytic syndrome in patients with TLR4 polymorphism.

TLR4 polymorphism produces impact on the efficiency of antiviral therapy of chronic hepatitis C. This suggests that therapeutic strategies should be adjusted not only to HCV genotype but also to individual TLR polymorphism.

Acknowledgments Igor Kajdashev, prof., the Pro-Rector for Research Efforts, HIGHER MEDICAL EDUCATIONAL INSTITUTION OF UKRAINE "UKRAINIAN MEDICAL STOMATOLOGICAL ACADEMY", Poltava, Ukraine

Toll-like receptor 4 polymorphism influence on the clinical course of chronic hepatitis C in HIV-infected patients RelevanceSlide Number 3ObjectivesSlide Number 5Description of a cohort of 62 patients infected with HIV/HCV and studied in relation to SNP of TLR4�(No. (%) of patients)Slide Number 7��������������������������Genotypic distribution of TLR4 SNP between healthy control subjects, HIV-positive patients, and HIV/HCV co-infected patients�Prevalence of Symptoms in persons with HIV/HCV co-infection in relation to SNP of the Toll-like receptor 4 gene �(No. (%) of patients)Slide Number 10���The development of hepatic fibrosis depending on TLR4 genotype variants������SVR of Chronic Hepatitis C therapy in HIV/HCV co-infected persons�ConclusionAcknowledgments