Supplementary Table 1. Laboratory reference ranges

Parameter / Reference
Haemoglobin (g/dl) / 11.0-18.0
ALT(IU/l) / 8-54
AST (IU/l) / 17-60
Creatinine (µmol/l) / 50-130
Hb1Ac (mmol/mol) / <42
CD4 count(cells/mm3) / 410-1590

Supplementary Table 2. HIV/HBV positive patients with significant tubular proteinuria (sTuPr)a

With sTuPr / Without sTuPr
N=9 / N=92
Female gender, n (%) / 8 (88.9) / 59 (64.1)
Age, median years (IQR) / 42 (38, 48) / 44 (39, 48)
BMI, median kg/m2(IQR) / 22.6 (20.3, 24.2) / 23.5 (20.5, 27.6)
Duration HIV diagnosis, median years(IQR) / 8.0 (5.3, 9.6) / 8.4 (6.7, 10.2)
ART duration, median years (IQR) / 6.0 (5.0, 7.9) / 8.0 (6.4, 9.2)
TDF duration, median years (IQR) / 4.0 (3.8, 4.1) / 4.0 (3.7, 4.1)
Prior ZDV, n (%) / 7 (77.8) / 82 (89.1)
Prior d4T, n (%) / 4 (44.4) / 44 (47.8)
Receiving EFV, n (%) / 8 (88.9) / 79 (85.9)
Receiving NVP, n (%) / 0 (0) / 4 (4.4)
Receiving LPV/r, n (%) / 1 (11.1) / 9 (9.8)
Adherence ≥90%, n (%) / 9 (100) / 77 (83.7)
Haemoglobin, median g/dl (IQR) / 11.6 (10.5, 12.0) / 13.4 (11.7, 14.7)
CD4 count, median cells/mm3 (IQR) / 463 (306, 628) / 573 (390, 717)
HIV-1 RNA >40 copies/ml, n (%) / 3 (33.3) / 18 (19.6)
HIV-1 RNA >1000 copies/ml, n (%) / 2 (22.2) / 12 (13.0)
HBV DNA >15 IU/ml, n (%) / 1 (11.1) / 14 (15.4)
HBV DNA >2000 IU/ml, n (%) / 0 (0) / 6 (6.5)
Liver stiffness, median kPa(IQR) / 4.8 (4.7, 8.8) / 4.5 (3.8, 5.8)
Liver stiffness kPa >9.4, n (%) / 2 (22.2) / 5 (5.4)
AST, median IU/l(IQR) / 26 (25, 37) / 31 (23, 39)
ALT, median IU/l(IQR) / 25 (14, 31) / 24 (18, 35)
HBeAgpositive, n (%) / 1 (9.1) / 10 (10.9)
Systolic BP, median mmHg(IQR) / 140 (122, 160) / 121 (110, 145)
Diastolic BP, median mmHg(IQR) / 90 (76, 110) / 80 (70, 90)
Hypertension, n (%) / Any grade / 5 (55.6) / 30 (32.6)
Grade 1b / 1 (11.1) / 12 (13.0)
Grade 2 / 1 (11.1) / 11 (12.0)
Grade 3 / 3 (33.3) / 7 (7.6)
NA / 0 (0) / 1 (1.1)
HB1Ac, median mmol/mol(IQR) / 33 (32, 38) / 33 (31, 38)
Diabetes, n (%)c / 2 (22.2) / 4 (4.5)
Schistosoma Ag positive, n (%) / 3 (33.3) / 15 (16.3)
Serum creatinine / median µmol/l (IQR) / 77 (55, 82) / 78 (67, 89)
eGFR / median ml/min (IQR) / 112 (81.2, 130) / 103 (92.6, 115)
≥90 ml/min / n (%) / 6 (66.7) / 73 (79.4)
60-89 ml/min / n (%) / 2 (22.2) / 16 (17.4)
30-60 ml/min / n (%) / 1 (11.1) / 3 (3.3)
change over time / median ml/min (IQR) / -2.77 (-7.93, -1.55) / -1.67 (-4.27, +0.29)
rapid decline (n=97) / n (%) / 3 (37.5) / 18 (20.2)
uPCR / median mg/mmol (IQR) / 53 (39, 58) / 13 (10, 19)
>20 mg/mmol / n (%) / 9 (100) / 19 (20.7)
≥50mg/mmol / n (%) / 5 (55.6) / 8 (8.7)
Dipstick protein / n (%) / 3 (33.3) / 12 (13.0)
Dipstick RBC / n (%) / 3 (33.3) / 9 (9.8)
Dipstick glucose / n (%) / 2 (22.2) / 15 (16.3)

aSignificant tubular proteinuria was defined by uPCR ≥30 mg/mmol and uACR/uPCR <0.4; bComprising 4 subjects with normal readings while on anti-hypertensive medication; cComprising 2 subjects with normal readings while on oral hypoglycaemic medication. IQR=inter-quartile range BMI=body mass index; ART=antiretroviral therapy; TDF=tenofovir disoproxil fumarate; ZDV=zidovudine; d4T=stavudine; EFV=efavirenz; NVP=nevirapine; LPV/r=ritonavir-boosted lopinavir; cps=copies; AST=aspartate transaminase; ALT=alanine transaminase; BP=blood pressure; NA=not available; Hb1Ac=glycated haemoglobin; Ag=Antigen; RD-eGFR=rapid declining estimated glomerular filtration rate (≥5 ml/min eGFR decline per year); uPCR=urinary protein/creatinine ratio; uACR=urinary albumin/creatinine ratio; RBC=red blood cells.

Supplementary Table 3. Additional multivariable logistic regression analyses of factors associated with TuPr in HIV/HBV positive patients with long-termTDF exposure

Model 1a / Model 2b / Model 3c
OR / 95% CI / p / OR / 95% CI / p / OR / 95% CI / p
Gender / female vs. male / 13.4 / 1.28, 139 / 0.03 / 11.5 / 1.30, 101 / 0.03 / 9.19 / 1.21, 75.4 / 0.04
Liver stiffness / per kPahigher / 1.09 / 0.94, 1.26 / 0.24 / - / - / - / - / - / -
AST / per 10 IU/l higher / - / - / - / 1.14 / 0.87, 1.50 / 0.34 / - / - / -
Hypertension / yes vs. no / 4.21 / 1.22, 14.5 / 0.02 / 4.26 / 1.30, 14.0 / 0.02 / 3.65 / 1.09, 12.3 / 0.04
HbA1c / per 5 mmol/mol higher / - / - / - / - / - / - / 1.07 / 0.88, 1.30 / 0.52
Schistosoma Ag / positive vs. negative / 1.18 / 0.26, 5.30 / 0.83 / 1.87 / 0.49, 7.14 / 0.36 / 1.95 / 0.51, 7.40 / 0.33

aModel 1 adjusted for gender, hypertension, Schistosoma Ag, and liver stiffness; bModel 2 adjusted for gender, hypertension, Schistosoma Ag, and AST level; cModel 3 adjusted for gender, hypertension, Schistosoma Ag, and HbA1c. OR=odds ratio; CI=confidence interval; AST=aspartate transaminase; Hb1Ac=glycated haemoglobin; Ag=antigen.

Supplementary Table 4. Urinary dipstick in relation to a diagnosis ofTuPr, RD-eGFR, hypertension, diabetes, and positive urinary Schistosoma antigen

TuPr / RD-eGFR / Hypertension / Diabetes / SchistosomaAg+
N=16 / N=21 / N=35 / N=6 / N=18
Haematuria / trace / 2 (12.5) / 0 (0) / 1 (2.9) / 0 (0) / 1 (5.6)
+ / 1 (6.3) / 1 (4.8) / 2 (5.7) / 0 (0) / 2 (11.1)
++ / 2 (12.5) / 2 (9.5) / 5 (14.3) / 1 (16.7) / 2 (11.1)
Glycosuria / trace / 3 (18.8) / 4 (19.1) / 8 (22.9) / 1 (16.7) / 4 (22.2)
+ / 0 (0) / 1 (4.8) / 2 (5.7) / 2 (33.3) / 1 (5.6)
++ / 1 (6.3) / 0 (0) / 1 (2.9) / 1 (16.7) / 0 (0)
Proteinuria / trace / 2 (12.5) / 1 (4.8) / 4 (11.4) / 1 (16.7) / 0 (0)
+ / 2 (12.5) / 1 (4.8) / 4 (11.4) / 0 (0) / 4 (22.2)
++ / 0 (0) / 1 (4.8) / 3 (8.6) / 1 (16.7) / 1 (5.6)

Haematuria (RBC/µL): Trace 5-10, + 50, ++ 250; glycosuria (mmol/L): Trace ≤1.1, + ≤2.8; ++ ≤ 55.5; proteinuria (g/L): Trace=≤0.3, +≤1, ++≤ 5. RBC=red blood cells; TuPr=tubular proteinuria; RD-eGFR=rapidly decliningestimated glomerular filtration rate; Ag=antigen; RBC=red blood cells.

Supplementary Figure 1. ROC analysis on the performance of dipstick proteinuria (a), dipstick glycosuria (b) and dipstick haematuria (c) as predictors of TuPr

a)b)

c)