Tuberculosis xxx (2015) 1e7
Contents lists available at ScienceDirect
Tuberculosis journal homepage: http://intl.elsevierhealth.com/journals/tube
IMMUNOLOGICAL ASPECTS
Cytokine and chemokine expression profiles in response to Mycobacterium tuberculosis stimulation are altered in HIV-infected compared to HIV-uninfected subjects with active tuberculosis Jillian L.M. Waruk a, *, Zipporah Machuki b, c, Christine Mesa a, Jennifer A. Juno b, Omu Anzala c, Meenu Sharma b, d, T. Blake Ball a, b, c, e, Julius Oyugi b, c, Sandra Kiazyk a, b, d a
National HIV Research Laboratory, Public Health Agency of Canada, JC Wilt Infectious Diseases Research Laboratory, Winnipeg, Canada Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada Department of Medical Microbiology, University of Nairobi, Nairobi, Kenya d National Reference Centre for Mycobacteriology, Public Health Agency of Canada, Winnipeg, Canada e Department of Immunology, University of Manitoba, Winnipeg, Canada b c
a r t i c l e i n f o
a b s t r a c t
Article history: Received 8 December 2014 Accepted 2 May 2015
Mycobacterium tuberculosis (Mtb) infects nearly 2 million people annually and is the most common cause of death in HIV-infected individuals. Tuberculosis (TB) diagnostics cater to HIV-uninfected individuals in non-endemic countries, are expensive, slow, and lack sensitivity for those most affected. Patterns of soluble immune markers from Mtb-stimulated immune cells are not well defined in HIV co-infection. We assessed immune differences between HIV-infected and HIV-uninfected individuals with active TB utilizing IFNg-based QuantiFERON®-TB Gold In-Tube (QFT) testing in Nairobi, Kenya. Excess QFT supernatants were used to measure cytokine and chemokine responses by a 17-plex bead array. Mtb/HIV coinfected participants were significantly less likely to be QFT+ (47.2% versus 84.2% in the HIV-uninfected group), and demonstrated lower expression of all cytokines except for IFNa2. Receiver operator characteristic analyses identified IL-1a as a potential marker of co-infection. Among HIV-infected individuals, CD4+ T cell count correlated weakly with the expression of several analytes. Co-expression analysis highlighted differences in immune profiles between the groups. These data suggest that there is a unique and detectable Mtb-specific immune response in co-infection. A better understanding of Mtb immunology can translate into much needed immunodiagnostics with enhanced sensitivity in HIV-infected individuals, facilitating their opportunity to obtain live-saving treatment. © 2015 Elsevier Ltd. All rights reserved.
Keywords: Tuberculosis HIV Co-infection Cytokine Chemokine Interferon gamma release assay ROC curve analysis
1. Introduction Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is the leading cause of death among individuals co-infected with HIV, most commonly affecting those with CD4 counts below 500 cells/mm3 [1e4]. HIV-infected individuals are at 20 to 37 times higher risk of reactivating from a latent TB infection (LTBI) to active TB disease (ATB) than the HIV-uninfected population [5,6]. More
Abbreviations: mtb, Mycobacterium tuberculosis; LTBI, latent TB infection; ATB, active TB; Treg, regulatory T cell; QFT, Quantiferon; IGRA, intereferon-gamma release assay. * Corresponding author. 1015 Arlington Street, Winnipeg, MB, R3E 3P6, Canada. Tel.: þ1 204 789 6485; fax: þ1 204 318 2222. E-mail address: [email protected] (J.L.M. Waruk).
than 95% of TB-related deaths occur in low- and middle-income TB endemic countries. The continent of Africa is especially hard-hit, with TB and HIV acting as co-epidemics [7]. At nearly 300 TB cases/100,000 population, Kenya is 10th among the top 22 high-burden countries in the world [7]. In Kenya and many sub-Saharan countries, pulmonary ATB is diagnosed by a combination of clinical observation of symptoms and sputum smear [8]. Mtb is only cultured on re-treatment or treatment failure for drug susceptibility. Tuberculin skin test (TST) is not commonly utilized due to low specificity for distinguishing patients with a history of TB illness, current infection, environmental Mycobacterin (BCG) vaccination [8]. rial exposure, and Bacille Calmette Gue The interferon gamma release assay (IGRA), which overcomes specificity issues in the TST by employing Mtb-specific antigens to elicit interferon gamma (IFNg) responses in whole blood, is utilized
http://dx.doi.org/10.1016/j.tube.2015.05.001 1472-9792/© 2015 Elsevier Ltd. All rights reserved.
Please cite this article in press as: Waruk JLM, et al., Cytokine and chemokine expression profiles in response to Mycobacterium tuberculosis stimulation are altered in HIV-infected compared to HIV-uninfected subjects with active tuberculosis, Tuberculosis (2015), http://dx.doi.org/ 10.1016/j.tube.2015.05.001
2
J.L.M. Waruk et al. / Tuberculosis xxx (2015) 1e7
in many developed countries, but its application in low- and middle-income countries is thought to be limited, especially in areas where HIV is a co-epidemic [9]. Immunodiagnosis of ATB in HIV-infected patients is imperfect due to immune dysfunction, and HIV-infected individuals can often present with extra-pulmonary TB or unusual symptoms. The protective immune response to Mtb infection has been long described as being TH1-like, whereby IFNg, IL-12, TNFa, and IP-10 dominate the cytokine and chemokine response [10,11]. In HIV infection, the depletion of CD4þ T cells severely impairs the host's response to other infectious diseases, especially in the case of Mtb, as HIV specifically targets Mtb-specific T cells for infection and destruction [12]. There have been a limited number of studies describing differential immune profiles between HIV-infected, Mtb-infected, and co-infected populations. These immune differences likely contribute to advanced rates of ATB in HIV co-infection [13,14]. Losses of cell types such as Th1/Th17 or regulatory CD4þ T cells (Treg) may contribute to the skewing of TB-specific immune responses away from an IFNg-dominated response in co-infected subjects [15,16]. Studies of soluble immune markers in HIV and Mtb co-infection are limited [17e19], with only two studies describing Mtb antigenspecific soluble immune responses [20,21]. Co-infected patients with ATB were shown to have significantly higher IL-10 expression levels that were independent of CD4 count, suggesting that HIVinfected individuals with active TB can mount detectable and distinguishable immune responses [20]. Interestingly, blockade of IL-10 receptor did not restore Mtb-specific IFNg, TNFa, IL-6, or IL-13 responses, thus the role of IL-10 has yet to be determined in coinfection. Another study demonstrated that plasma IL-10 levels were not different between HIV-infected and HIV-uninfected participants with ATB, though after Mtb antigen stimulation, significantly higher levels of IL-10 and IL-2, but not IFNg or IL-18, were observed in HIV-uninfected and HIV-infected participants with CD4 counts >200 cell/mm3 [21]. There are no clear trends observed in the comparison of HIV-infected and HIV-uninfected people with active TB beyond a general immune suppression in co-infection, and antigen-specific soluble responses are only minimally described utilizing few read-outs. This study aims to better define Mtb antigen-specific immunological disparities that exist between HIV-infected and HIVuninfected people with active TB disease in an attempt to inform both TB immunodiagnostics and vaccine approaches towards optimal responses measurable in the HIV/Mtb co-infected population in an endemic setting. 2. Materials and methods 2.1. Study subjects Study participants were enrolled within the Respiratory Clinic at the Mbagathi Provincial Hospital in Nairobi, Kenya. Patients attend the hospital primarily for TB screening and treatment. In addition, we also enrolled household contacts accompanying the ATB infected participants to the clinic to utilize as a comparison group. All study participants gave informed consent, and the University of Nairobi Research Ethics Board approved ethics for the study. All participants were administered the QuantiFERON®-TB Gold InTube (QFT) IFNg-based TB test. Patients suspected of having an active pulmonary TB infection were submitted to a confirmatory chest x-ray and/or their sputum to acid-fast bacilli sputum smear test microscopy. Only patients with at least two confirmatory positive tests (chest x-ray, sputum smear, QFT) for ATB were included in this study and any patient with unconfirmed diagnosis (i.e. negative for any two of the chest x-ray, smear, or QFT test) were
removed from the study. Only participants agreeing to HIV testing were enrolled. When possible, the results of x-ray, smear microscopy, ARV status, and most recent CD4 count were recorded. HIV testing was done according to Government of Kenya guidelines; the Alere Determine™ HIV-1/2 Ag/Ab Combo test and SD Bioline HIV1/2 test (Standard Diagnostics, Inc., Gyeonggi-do, Republic of Korea) were used for diagnosis and positive result confirmation, respectively. In the case of test disagreement, the Uni-Gold™ Recombigen® HIV1/2 test (Trinity Biotech, Wicklow, Ireland) was employed as a tiebreaker. Due to low numbers of enrolment, HIVinfected contacts were excluded from the study. In total, 53 HIVinfected participants with active TB and 57 HIV-uninfected participants with active TB were enrolled. Thirteen HIV negative, TB negative household contacts were enrolled.
2.2. Interferon-gamma release assay The QuantiFERON®-TB Gold In-Tube test (Qiagen, USA) was performed according to the manufacturer's instructions. Briefly, blood was collected from the participant into 3 blood collection tubes e one containing Mtb peptide antigens ESAT-6, CFP-10, and TB7.7, one containing a mitogen, and one nil tube, with no antigen. Whole blood was incubated for 16 h at 37 C and transferred to 4 C until processing, but for no longer than 48 h. Tubes were centrifuged at 2000 g for 15 min, and then supernatants were stored at 80 C until the QFT ELISA could be conducted. Supernatants and IFNg standards (50 ml) plus conjugate (50 ml) were incubated for 2 h, washed 6 times, and incubated for 30 min with substrate solution. After 30 min, 50 ml stop solution was added and the wells and the plates were read at 450 nm with a 650 nm reference filter. Concentrations of IFNg were calculated based on the standard curve. Test outcomes of positive, negative, or indeterminate were determined according to the manufacturer's mathematical algorithm. All participants with indeterminate QFT results had their samples rerun and positive or negative outcomes on second run were used for their final results. Those still indeterminate on second run were labelled as QFT indeterminate. The remainder of the QFT supernatants was used for the milliplex assays.
2.3. Multiplexed cytokine and chemokine arrays Milliplex assays were performed using the MILLIPLEX® MAP Human Cytokine/Chemokine Magnetic Bead Panel Immunology Multiplex Assay (Merck Milliplore, Billerica, USA) designed to measure interferon (IFN) a2, IFNg, interleukin (IL)-1a, IL-1b, IL-1 receptor antagonist (RA), IL-2, IL-6, IL-8, IL-10, IL-12 P40, IL-12 P70, IL-15, IL-17A, IFNg-induced protein (IP)-10, macrophage inflammatory protein (MIP)-1a, MIP-1b, and tumour necrosis factor (TNF)a. Briefly, beads coated with antibodies against the 17 cytokines and chemokines were added to a 96-well filter plate in duplicate along with the neat samples and standard serial dilutions plus serum matrix. The plate was incubated for 2 h at room temperature, washed and detection antibody was added to the plate. After 1 h at room temperature, the plate was further incubated for 30 min with detection streptavidin-phycoerythrin. The plate was washed and 150 ml of sheath fluid was added for analysis on the Luminex 100™ IS (Luminex, Austin, TX). The median fluorescent intensity (MFI) data was analysed using weighted 5-parameter logistic for calculating the cytokine and chemokine concentrations in the samples based on the standard curves. Cytokine responses to antigen were calculated as the difference in the mean measured cytokine of the background expression levels in the nil plasma from the TB antigen plasma.
Please cite this article in press as: Waruk JLM, et al., Cytokine and chemokine expression profiles in response to Mycobacterium tuberculosis stimulation are altered in HIV-infected compared to HIV-uninfected subjects with active tuberculosis, Tuberculosis (2015), http://dx.doi.org/ 10.1016/j.tube.2015.05.001
J.L.M. Waruk et al. / Tuberculosis xxx (2015) 1e7
3
Table 1 Study participant characteristics. Study subject characteristic
ATB* HIVþ
ATB HIV
HIV contacts
n Sex Male (%)z Female (%) No data (%)x Mean age (IQR)k QFT þ (%) QFT - (%) QFT indeterminate (%) QFT no data (%) Chest x-ray þ (%) Chest x-ray e (%) Chest x-ray no data (%) Microscopy þ (%) Microscopy e (%) Microscopy no data (%) CD4 counts > 200 (range) < 200 (range)
53
57
13
22 (41.5) 30 (56.7) 1 (1.9) 36 (29e45) 25 (47.2) 19 (35.9) 9 (17.0) 0 (0) 35 (100) 0 (0) 18 (34.0) 19 (90.5) 2 (9.5) 32 (60.4) 19 (217e695) 18 (2e175)
35 (62.7) 22 (37.3)
1 (12.5) 12 (87.5)
0.0561¶
32 (24e35) 48 (84.2) 9 (15.8) 0 (0) 0 (0) 43 (100) 0 (0) 16 (28.1) 26 (72.2) 10 (27.8) 21 (36.8) No data
35 (25e46) 0 (0) 13 (100) 0 (0) 0 (0) No data
0.0015¶
Contents lists available at ScienceDirect
Tuberculosis journal homepage: http://intl.elsevierhealth.com/journals/tube
IMMUNOLOGICAL ASPECTS
Cytokine and chemokine expression profiles in response to Mycobacterium tuberculosis stimulation are altered in HIV-infected compared to HIV-uninfected subjects with active tuberculosis Jillian L.M. Waruk a, *, Zipporah Machuki b, c, Christine Mesa a, Jennifer A. Juno b, Omu Anzala c, Meenu Sharma b, d, T. Blake Ball a, b, c, e, Julius Oyugi b, c, Sandra Kiazyk a, b, d a
National HIV Research Laboratory, Public Health Agency of Canada, JC Wilt Infectious Diseases Research Laboratory, Winnipeg, Canada Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada Department of Medical Microbiology, University of Nairobi, Nairobi, Kenya d National Reference Centre for Mycobacteriology, Public Health Agency of Canada, Winnipeg, Canada e Department of Immunology, University of Manitoba, Winnipeg, Canada b c
a r t i c l e i n f o
a b s t r a c t
Article history: Received 8 December 2014 Accepted 2 May 2015
Mycobacterium tuberculosis (Mtb) infects nearly 2 million people annually and is the most common cause of death in HIV-infected individuals. Tuberculosis (TB) diagnostics cater to HIV-uninfected individuals in non-endemic countries, are expensive, slow, and lack sensitivity for those most affected. Patterns of soluble immune markers from Mtb-stimulated immune cells are not well defined in HIV co-infection. We assessed immune differences between HIV-infected and HIV-uninfected individuals with active TB utilizing IFNg-based QuantiFERON®-TB Gold In-Tube (QFT) testing in Nairobi, Kenya. Excess QFT supernatants were used to measure cytokine and chemokine responses by a 17-plex bead array. Mtb/HIV coinfected participants were significantly less likely to be QFT+ (47.2% versus 84.2% in the HIV-uninfected group), and demonstrated lower expression of all cytokines except for IFNa2. Receiver operator characteristic analyses identified IL-1a as a potential marker of co-infection. Among HIV-infected individuals, CD4+ T cell count correlated weakly with the expression of several analytes. Co-expression analysis highlighted differences in immune profiles between the groups. These data suggest that there is a unique and detectable Mtb-specific immune response in co-infection. A better understanding of Mtb immunology can translate into much needed immunodiagnostics with enhanced sensitivity in HIV-infected individuals, facilitating their opportunity to obtain live-saving treatment. © 2015 Elsevier Ltd. All rights reserved.
Keywords: Tuberculosis HIV Co-infection Cytokine Chemokine Interferon gamma release assay ROC curve analysis
1. Introduction Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), is the leading cause of death among individuals co-infected with HIV, most commonly affecting those with CD4 counts below 500 cells/mm3 [1e4]. HIV-infected individuals are at 20 to 37 times higher risk of reactivating from a latent TB infection (LTBI) to active TB disease (ATB) than the HIV-uninfected population [5,6]. More
Abbreviations: mtb, Mycobacterium tuberculosis; LTBI, latent TB infection; ATB, active TB; Treg, regulatory T cell; QFT, Quantiferon; IGRA, intereferon-gamma release assay. * Corresponding author. 1015 Arlington Street, Winnipeg, MB, R3E 3P6, Canada. Tel.: þ1 204 789 6485; fax: þ1 204 318 2222. E-mail address: [email protected] (J.L.M. Waruk).
than 95% of TB-related deaths occur in low- and middle-income TB endemic countries. The continent of Africa is especially hard-hit, with TB and HIV acting as co-epidemics [7]. At nearly 300 TB cases/100,000 population, Kenya is 10th among the top 22 high-burden countries in the world [7]. In Kenya and many sub-Saharan countries, pulmonary ATB is diagnosed by a combination of clinical observation of symptoms and sputum smear [8]. Mtb is only cultured on re-treatment or treatment failure for drug susceptibility. Tuberculin skin test (TST) is not commonly utilized due to low specificity for distinguishing patients with a history of TB illness, current infection, environmental Mycobacterin (BCG) vaccination [8]. rial exposure, and Bacille Calmette Gue The interferon gamma release assay (IGRA), which overcomes specificity issues in the TST by employing Mtb-specific antigens to elicit interferon gamma (IFNg) responses in whole blood, is utilized
http://dx.doi.org/10.1016/j.tube.2015.05.001 1472-9792/© 2015 Elsevier Ltd. All rights reserved.
Please cite this article in press as: Waruk JLM, et al., Cytokine and chemokine expression profiles in response to Mycobacterium tuberculosis stimulation are altered in HIV-infected compared to HIV-uninfected subjects with active tuberculosis, Tuberculosis (2015), http://dx.doi.org/ 10.1016/j.tube.2015.05.001
2
J.L.M. Waruk et al. / Tuberculosis xxx (2015) 1e7
in many developed countries, but its application in low- and middle-income countries is thought to be limited, especially in areas where HIV is a co-epidemic [9]. Immunodiagnosis of ATB in HIV-infected patients is imperfect due to immune dysfunction, and HIV-infected individuals can often present with extra-pulmonary TB or unusual symptoms. The protective immune response to Mtb infection has been long described as being TH1-like, whereby IFNg, IL-12, TNFa, and IP-10 dominate the cytokine and chemokine response [10,11]. In HIV infection, the depletion of CD4þ T cells severely impairs the host's response to other infectious diseases, especially in the case of Mtb, as HIV specifically targets Mtb-specific T cells for infection and destruction [12]. There have been a limited number of studies describing differential immune profiles between HIV-infected, Mtb-infected, and co-infected populations. These immune differences likely contribute to advanced rates of ATB in HIV co-infection [13,14]. Losses of cell types such as Th1/Th17 or regulatory CD4þ T cells (Treg) may contribute to the skewing of TB-specific immune responses away from an IFNg-dominated response in co-infected subjects [15,16]. Studies of soluble immune markers in HIV and Mtb co-infection are limited [17e19], with only two studies describing Mtb antigenspecific soluble immune responses [20,21]. Co-infected patients with ATB were shown to have significantly higher IL-10 expression levels that were independent of CD4 count, suggesting that HIVinfected individuals with active TB can mount detectable and distinguishable immune responses [20]. Interestingly, blockade of IL-10 receptor did not restore Mtb-specific IFNg, TNFa, IL-6, or IL-13 responses, thus the role of IL-10 has yet to be determined in coinfection. Another study demonstrated that plasma IL-10 levels were not different between HIV-infected and HIV-uninfected participants with ATB, though after Mtb antigen stimulation, significantly higher levels of IL-10 and IL-2, but not IFNg or IL-18, were observed in HIV-uninfected and HIV-infected participants with CD4 counts >200 cell/mm3 [21]. There are no clear trends observed in the comparison of HIV-infected and HIV-uninfected people with active TB beyond a general immune suppression in co-infection, and antigen-specific soluble responses are only minimally described utilizing few read-outs. This study aims to better define Mtb antigen-specific immunological disparities that exist between HIV-infected and HIVuninfected people with active TB disease in an attempt to inform both TB immunodiagnostics and vaccine approaches towards optimal responses measurable in the HIV/Mtb co-infected population in an endemic setting. 2. Materials and methods 2.1. Study subjects Study participants were enrolled within the Respiratory Clinic at the Mbagathi Provincial Hospital in Nairobi, Kenya. Patients attend the hospital primarily for TB screening and treatment. In addition, we also enrolled household contacts accompanying the ATB infected participants to the clinic to utilize as a comparison group. All study participants gave informed consent, and the University of Nairobi Research Ethics Board approved ethics for the study. All participants were administered the QuantiFERON®-TB Gold InTube (QFT) IFNg-based TB test. Patients suspected of having an active pulmonary TB infection were submitted to a confirmatory chest x-ray and/or their sputum to acid-fast bacilli sputum smear test microscopy. Only patients with at least two confirmatory positive tests (chest x-ray, sputum smear, QFT) for ATB were included in this study and any patient with unconfirmed diagnosis (i.e. negative for any two of the chest x-ray, smear, or QFT test) were
removed from the study. Only participants agreeing to HIV testing were enrolled. When possible, the results of x-ray, smear microscopy, ARV status, and most recent CD4 count were recorded. HIV testing was done according to Government of Kenya guidelines; the Alere Determine™ HIV-1/2 Ag/Ab Combo test and SD Bioline HIV1/2 test (Standard Diagnostics, Inc., Gyeonggi-do, Republic of Korea) were used for diagnosis and positive result confirmation, respectively. In the case of test disagreement, the Uni-Gold™ Recombigen® HIV1/2 test (Trinity Biotech, Wicklow, Ireland) was employed as a tiebreaker. Due to low numbers of enrolment, HIVinfected contacts were excluded from the study. In total, 53 HIVinfected participants with active TB and 57 HIV-uninfected participants with active TB were enrolled. Thirteen HIV negative, TB negative household contacts were enrolled.
2.2. Interferon-gamma release assay The QuantiFERON®-TB Gold In-Tube test (Qiagen, USA) was performed according to the manufacturer's instructions. Briefly, blood was collected from the participant into 3 blood collection tubes e one containing Mtb peptide antigens ESAT-6, CFP-10, and TB7.7, one containing a mitogen, and one nil tube, with no antigen. Whole blood was incubated for 16 h at 37 C and transferred to 4 C until processing, but for no longer than 48 h. Tubes were centrifuged at 2000 g for 15 min, and then supernatants were stored at 80 C until the QFT ELISA could be conducted. Supernatants and IFNg standards (50 ml) plus conjugate (50 ml) were incubated for 2 h, washed 6 times, and incubated for 30 min with substrate solution. After 30 min, 50 ml stop solution was added and the wells and the plates were read at 450 nm with a 650 nm reference filter. Concentrations of IFNg were calculated based on the standard curve. Test outcomes of positive, negative, or indeterminate were determined according to the manufacturer's mathematical algorithm. All participants with indeterminate QFT results had their samples rerun and positive or negative outcomes on second run were used for their final results. Those still indeterminate on second run were labelled as QFT indeterminate. The remainder of the QFT supernatants was used for the milliplex assays.
2.3. Multiplexed cytokine and chemokine arrays Milliplex assays were performed using the MILLIPLEX® MAP Human Cytokine/Chemokine Magnetic Bead Panel Immunology Multiplex Assay (Merck Milliplore, Billerica, USA) designed to measure interferon (IFN) a2, IFNg, interleukin (IL)-1a, IL-1b, IL-1 receptor antagonist (RA), IL-2, IL-6, IL-8, IL-10, IL-12 P40, IL-12 P70, IL-15, IL-17A, IFNg-induced protein (IP)-10, macrophage inflammatory protein (MIP)-1a, MIP-1b, and tumour necrosis factor (TNF)a. Briefly, beads coated with antibodies against the 17 cytokines and chemokines were added to a 96-well filter plate in duplicate along with the neat samples and standard serial dilutions plus serum matrix. The plate was incubated for 2 h at room temperature, washed and detection antibody was added to the plate. After 1 h at room temperature, the plate was further incubated for 30 min with detection streptavidin-phycoerythrin. The plate was washed and 150 ml of sheath fluid was added for analysis on the Luminex 100™ IS (Luminex, Austin, TX). The median fluorescent intensity (MFI) data was analysed using weighted 5-parameter logistic for calculating the cytokine and chemokine concentrations in the samples based on the standard curves. Cytokine responses to antigen were calculated as the difference in the mean measured cytokine of the background expression levels in the nil plasma from the TB antigen plasma.
Please cite this article in press as: Waruk JLM, et al., Cytokine and chemokine expression profiles in response to Mycobacterium tuberculosis stimulation are altered in HIV-infected compared to HIV-uninfected subjects with active tuberculosis, Tuberculosis (2015), http://dx.doi.org/ 10.1016/j.tube.2015.05.001
J.L.M. Waruk et al. / Tuberculosis xxx (2015) 1e7
3
Table 1 Study participant characteristics. Study subject characteristic
ATB* HIVþ
ATB HIV
HIV contacts
n Sex Male (%)z Female (%) No data (%)x Mean age (IQR)k QFT þ (%) QFT - (%) QFT indeterminate (%) QFT no data (%) Chest x-ray þ (%) Chest x-ray e (%) Chest x-ray no data (%) Microscopy þ (%) Microscopy e (%) Microscopy no data (%) CD4 counts > 200 (range) < 200 (range)
53
57
13
22 (41.5) 30 (56.7) 1 (1.9) 36 (29e45) 25 (47.2) 19 (35.9) 9 (17.0) 0 (0) 35 (100) 0 (0) 18 (34.0) 19 (90.5) 2 (9.5) 32 (60.4) 19 (217e695) 18 (2e175)
35 (62.7) 22 (37.3)
1 (12.5) 12 (87.5)
0.0561¶
32 (24e35) 48 (84.2) 9 (15.8) 0 (0) 0 (0) 43 (100) 0 (0) 16 (28.1) 26 (72.2) 10 (27.8) 21 (36.8) No data
35 (25e46) 0 (0) 13 (100) 0 (0) 0 (0) No data
0.0015¶
