Institutes:Ukrainian Medical and Dental Academy,

Regional HIV/AIDS Prevention and Control Center, Poltava Ukraine

Toll-like receptor 4 polymorphism influence on the development of opportunistic diseases

in HIV-infected patients

The presenting author is Tetiana Kyrychenko

Authors: G. Dubynska, T.Koval, I.Kaidashev, V.Korshenko

Ukraine

11,012,5

14,817,3 18,0

28,3

34,237,8

40,943,2

44,746,6

0,0

5,0

10,0

15,0

20,0

25,0

30,0

35,0

40,0

45,0

50,0

2000. 2001. 2002. 2003. 2004. 2005. 2006. 2007. 2008. 2009. 2010. 2011.

Ukraine has one of the fastest growing HIV/AIDS 

epidemics in the world and the

highest in all of Europe. Late 2011 Ukraine

numbered 210,000 HIV-positive persons.

Dynamics of officially registered new cases of HIV-infection in Ukraine (on 100 thousand population)

Toll-like receptors (TLRs) have a key role in innate immunity and mutations in the genes encoding these

receptors have been associated with increased or decreased susceptibility to infections.

Objectives:

to study the prevalence of TLR4 Asp299Gly SNP among the healthy and HIV-positive persons in Ukraine;

to find out HIV clinical manifestations depending on TLR4 genotype variants.

Asp299Gly, A>G

chromosome 9q32–33

Methods Patients. The study included 180

antiretroviral treatment-naive HIV-1 infected patients.

The stage of the disease was recorded in accordance with the expanded European AIDS case definition.

Laboratory procedures. Routine CD-4 cell count was determined

using conventional flow cytometry with fresh blood samples from all patients.

Plasma HIV-RNA was also routinely measured by the polymerase chain reaction (PCR) (Real time HIV-1, Abbout).

TLR4 genotyping was performed by real-time PCR using oligonucleotide primers (Tercik PCR system, DNA-technology, Moscow)

Characteristic of a Cohort of 180 patients infected with HIV-1 and studied in Relation to

SNP of TLR4 Characteristic Wild type

(AA, n=165)

Asp299Gly heterozygous

(AG, n=15)

Male-to-female ratio 98:67 10:5

Ageª, mean years± standard deviation

35,21±0,51 35,13±1,33

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

259,52±16,2 233,6±30,38

Maximum viral loadª, median copies/mL

249826 152671

Clinical stage(%)ABC

11 (6,6)72 (43,7)82 (49,7)

2(13,3)2(13,3)11(73,4)

History of opportunistic infections in 180 HIV-infected patients

1%

1%

1%

4%

14%

18%

20%

25%

27%

28%

33%

51%

48%

9%

0% 10% 20% 30% 40% 50% 60%

Progressive multifocal leukoencephalopathy

Kaposi sarcoma

Pneumocystis jiroveci 

Cytomegalovirus disease

Toxoplasmosis of brain

TB extra pulmonary

Herpes simplex: chronic ulcers

Herpes zoster

Tuberculosis pulmonary

Wasting syndrome caused by HIV  

Bacterial pneumonia

Hairy leukoplakia, oral

Candidiasis

Chronic hepatitis C

History of opportunistic infections in 180 HIV-infected patients in relation to SNP of the Toll-like receptor 4 gene

(No. (%) of patients)

Variable Wild type (AA,n=165)

Asp299Gly heterozygous

(AG, n=15)

p

Mycobacterium tuberculosis, pulmonary or extrapulm.

59 (35.7) 11 (73.3) 0.007*

Bacterial pneumonia, recurrent

42 (25.5) 8 (53.3) 0.021*

Hairy leukoplakia, oral

51 (30.9) 9 (60) 0.022*

Herpes zoster (shingles), involving two or more episodes or at least one dermatome

29 (17.6) 7 (46.7) 0.007*

*AG compared to AA, Mann-Whitney U-test

History of opportunistic infections in 180 HIV-infected patients in relation to SNP of the Toll-like receptor 4 gene

(No. (%) of patients)

Variable Wild type (AA,n=165)

Asp299Gly heterozygous

(AG, n=15)

p

Chronic hepatitis C 80 (48.5) 12 (80) 0.018*

Candidiasis, oropharyngeal or esophageal

78 (47.3) 8 (53.3) >0.05

Wasting syndrome caused by HIV  (involuntary weight loss >10% of baseline body weight) associated with either chronic diarrhea or chronic weakness and documented fever for ≥1 month)

42 (25.5) 6 (40) >0.05

Herpes simplex: chronic ulcers (>1 month in duration)

28 (17) 5 (33.3) >0.05

Any 19 (11.5) 3 (20) >0.05

*AG compared to AA, Mann-Whitney U-test

Distribution of the 299 alleles of the patients when they were grouped by CD-4 T-cell counts equal to or above and below

350, 200 100

cells/mm³, respectively

*Only patients with CD4 T-cell

counts above 200cells/mm³ had the prevalence of the Asp299Gly polymorphism (12%) significantly higher than patients with CD4 T-cell counts below than 200 cells/mm³ (3.7%), p<0.05

133

1232

30

20

40

60

80

100

120

140

Nu

mb

er

of

ind

ivid

uals

≤350 >350

CD4 T cell/mm

AA

AG

77

3

88

12*

0102030405060708090

Nu

mb

er

of

ind

ivid

uals

≤200 >200

CD4 T cells/mm

AA

AG

312

134

13

0

20

40

60

80

100

120

140

Nu

mb

er

of

ind

ivid

uals

≤100 >100

CD-4 T cells/mm

AA

AG

Detection of opportunistic infections in HIV-infected patients with different levels of CD4

(No. (%) of patients)CD-4 T-cell counts /mm³

<100 101-349 >350

TLR-4 genotype

AA,n=31

AG, n=3

AA,n=102

AG, n=9

AA,n=32

AG, n=3

Tuberculosis 20(64.5) 3(100) 34(33.3) 6(66.7)* 5(15.6) 2(66.7)*

Toxoplasmosis 12(38.8) 3(100)* 0(0) 1(11.1)* 0 0

Herpes zoster 2(6.5) 1(33.3) 24(23.6) 5(55.6)* 3(9.3) 1(33.3)

Pneumonia 10(32.3) 1(33.3) 22(21.6) 5(55.6)* 10(31) 2(66.7)

*AG compared to AA, X²-test, p<0.05

Conclusions This study suggests a grater risk for

development of active tuberculosis, bacterial pneumonia, herpes zoster, hairy leukoplakia, toxoplasmosis of brain, chronic hepatitis C in patients with the Asp299Gly TLR4 polymorphism

our results demonstrate the impact of a TLR4 genetic polymorphism on development of opportunistic infections for CD4 levels more than 100.

Acknowledgments

G.Dubynska, prof., department of infectious diseases, Ukrainian Medical and Dental Academy, Poltava, Ukraine

T.Koval, prof., department of infectious diseases, Ukrainian Medical and Dental Academy, Poltava, Ukraine

I. Kajdashev, prof., chair of research department, Ukrainian Medical and Dental Academy, Poltava, Ukraine

V.Korshenko, chief doctor of Regional centre of HIV/AIDS Poltava, Ukraine