Table S1. Semen parameters of patients and controls undergoing sperm proteome analysis
HIV-positive patients, n=15,
median (range)
Controls, n=15
median (range)
pb
42 (26 - 49)
37 (27 - 58)
0.595
TDF/FTC/EFV
3
n.a.
n.a.
TDF/FTC/RLP
1
n.a.
n.a.
RAL/TDF/FTC
2
n.a.
n.a.
DRV/RTV/TDF/FTC
2
n.a.
n.a.
LPV/RTV/TDF/FTC/ABC
1
n.a.
n.a.
ATV/RTV/TDF/FTC
1
n.a.
n.a.
fAPV/RTV/TDF/FTC
1
n.a.
n.a.
NVP/TDF/FTC
2
n.a.
n.a.
NVP/ABC/3TC
2
n.a.
n.a.
4 (2.4 - 6.0)
3.3 (1.7 - 7.1)
0.116
Parameter
Patient age [years]
ART therapy
Semen volume [ml]
Semen pH
Sperm concentration [106/ml]
Progressive motility [%]
a
Normal morphology [%]
6
7.8 (7.3 - 8.5)
7.7 (7.3 - 8.3)
0.285
71.0 (25.8 - 425.0)
58.0 (19.4 - 194.0)
0.653
52 (25 - 77)
52 (9 - 72)
0.806
3.0 (0 - 9)
7.0 (2 - 24)
0.013
Peroxidase -positive leukocytes [10 /ml]
0.1 (0 - 0.6)
0.1 (0 - 8.9)
0.683
Seminal elastase [ng/ml]
84 (10 - 965)
68 (13 - 2000)
0.775
1
1
n.a.
75 (35 - 97)
79 (50 - 92)
0.806
6 c
Spermatozoa for proteome analysis [10 ]
Progressive motility in swim-up samples [%]
a
using strict criteria, bMann-Whitney U test, c1×106 sperms were taken from each patient to standardize protein concentration, n.a. not
applicable
Table S2. Characteristics of the study populationa
Parameter
Age (years)
Patients without comorbitities
(n=88)
Patients with comorbitities (n=28)
43 (18 - 64)
47 (24 - 61)
Duration of HIV infection (years)
5 (0 - 33)
8.5 (0 - 28)
Time on ART (years)
4 (0 - 20)
7 (0 - 17)
3
59 (67.0%)
16 (57.1%)
4
23 (26.1%)
10 (35.7%)
5
6 (6.8%)
2 (7.1%)
548.5 (23 - 1536)
449.5 (98 - 1151)
81 (92%)
25 (89.3%)
7 (8%)
3 (10.7%)
80212 (7542 - 6000000)
38371 (919 - 750000)
233 (3 - 762)
138.5 (4 - 461)
A
24 (27.3%)
5 (17.9%)
B
27 (30.7%)
10 (35.7%)
C
37 (42%)
13 (60.7%)
1
5 (5.7)
1 (3.6%)
2
43 (48.9)
10 (35.7%)
3
40 (45.5)
17 (60.7%)
heterosexual
30 (34.1%)
10 (35.7%)
homosexual
45 (51.1%)
16 (57.1%)
bisexual
13 (14.8%)
2 (7.1%)
49 (55.7%)
14 (50%)
50 (1 - 5000)
50 (5 - 1500)
23 (26.1%)
5 (17.9%)
FSH (mU/ml)
5.6 (1.5 - 23.4)
8.2 (0.6 - 72.5)
LH (mU/ml)
5.5 (2 - 30.1)
4.6 (0.1 - 47.1)
Testosterone (ng/dl)
505.5 (213 - 1006)
442 (79 - 1020)
Free Testosterone (ng/dl)
8.1 (3.15 - 20.23)
7.9 (0.9 - 14.7)
Prolactin (uIU/ml)
147 (59 - 1206)
133.5 (34 - 301)
Oestradiol (pg/ml)
31 (12 - 66)
32 (12-50)
43.3 (19.5 - 124)
47.5 (21.7 - 99.5)
12.7 (4.6 - 25.6)
12.9 (3.4 -22.3)
current
38 (43.2%)
13 (46.4%)
previous
12 (13.6%)
6 (21.4%)
Total
50 (56.8%)
18 (64.3%)
never
21 (23.9%)
4 (14.3%)
scarce
26 (29.5%)
11 (39.3%)
Number of current ART drugs
Present CD4 count (cells/µl)
Viral load in blood
below detection limit
detectable
Highest known viral load (copies/ml)
CD4 nadir (cells/µl)
CDC stage
Sexual orientation
Current partner
Number of lifetime sexual partners
Patients with own children
Hormonesb
SHBG (nmol/l)
Mean testicular volume (ml)
Smokers
Alcohol consumption
occasional
26 (29.5%)
9 (32.1%)
often
11 (12.5%)
2 (7.1%)
daily
4 (4.5%)
2 (7.1%)
current
12 (13.6%)
5 (17.9%)
previous
21 (23.9%)
6 (21.4%)
Total
26 (29.5%)
7 (25.0%)
Drug abuse
a
data are expressed either as median (range) or, when categorical, as absolute numbers and percentages
b
measured between 8 and 10 a.m.
Table S3. Correlation between semen parameters and markers of HIV disease
CD4 cell
count (n=88)
CDC Status (n=88)
Duration of
disease (n=88)
Duration of ART
(n=88)
Positive viral
loada (n=88)
ABC
123
r, p
r, p
r, p
r, p
r, p
p
Volume (ml)
NS
NS
NS
NS
NS
NS
pH value
NS
NS
r=0.239, p=0.026
NS
NS
NS
NS
NS
NS
NS
NS
NS
Total sperm count (10 )
NS
NS
NS
NS
NS
NS
Progressive motility (%)
NS
NS
NS
NS
NS
NS
Sperm vitality (%)
NS
NS
NS
NS
r=-0.228, p=0.037
NS
Normal morphology (%)
NS
NS
NS
NS
NS
NS
Alpha-glucosidase (mU/ejaculate)
NS
NS
NS
NS
NS
NS
Fructose (µmol/ejaculate)
NS
NS
NS
NS
NS
NS
Zinc (µmol/ejaculate)
NS
NS
NS
NS
NS
NS
Peroxidase+ leukocytes (10 /ml)
NS
NS
NS
NS
r=0.228, p=0.033
NS
Elastase (ng/ml)
NS
r=0.239, p=0.038
NS
NS
NS
NS
6
Sperm concentration (10 /ml)
6
6
a
Mann-Whitney U test, NS not significant, bscore calculated according to [23]
N
Table S4. Sperm protein alterations in HIV-positive patientsa
Protein name
Protein
accession
number
MW
[kDa]
Peptides
Sequence
coverage
[%]
Mascot
score
Regulation fact
patients/control
3523 FRMD4A
FERM domain-containing protein 4A
IPI00514952
60.2
6
16.8
62.4
0.01
3526 ACPP
Prostatic acid phosphatase
IPI00396434
44.5
11
26.7
102.0
0.35
3617 KRT10
Keratin, type I cytoskeletal 10
IPI00009865
58.8
17
29.3
116.0
0.16
3619 HSPA1A
Heat shock 70 kDa protein 1A/1B
IPI00304925
70.0
11
22.0
69.3
0.06
3622 HSPA1A
Heat shock 70 kDa protein 1A/1B
IPI00304925
70.0
9
18.7
64.6
0.08
4305 HSPB1
Heat shock protein beta-1
IPI00025512
22.8
9
59.0
109.0
0.32
4406 ANXA3
Annexin A3
IPI00024095
36.4
16
48.3
164.0
0.02
CKB
Creatine kinase B-type
IPI00022977
42.6
8
27.6
70.1
CKB
Creatine kinase B-type
IPI00908811
38.7
7
26.6
73.7
Serum albumin
IPI00022434
71.7
13
22.6
85.1
KRT10
Keratin, type I cytoskeletal 10
IPI00009865
58.8
13
21.6
83.3
KRT9
Keratin, type I cytoskeletal 9
IPI00019359
62.0
13
23.4
92.5
5301 PRDX6
Peroxiredoxin-6
IPI00220301
25.0
8
41.1
83.7
0.02
5402 KRT10
Keratin, type I cytoskeletal 10
IPI00009865
58.8
9
19.2
65.3
0.48
5502 KRT10
Keratin, type I cytoskeletal 10
IPI00009865
58.8
10
16.4
86.0
0.14
5508 KRT10
Keratin, type I cytoskeletal 10
IPI00009865
58.8
10
19.2
76.4
0.29
KRT10
Keratin, type I cytoskeletal 10
IPI00009865
58.8
19
30.0
135.0
KRT1
Keratin, type II cytoskeletal 1
IPI00220327
66.0
17
25.9
90.2
KRT10
Spot name
4502
Gene name
4608 ALB
4610
5610
a
0.24
0.28
0.16
0.18
Keratin, type I cytoskeletal 10
IPI00009865
58.8
17
30.5
122.0
5705 KRT6C
Keratin, type II cytoskeletal 6C
IPI00299145
60.0
14
24.5
80.9
KRT6C
Keratin, type II cytoskeletal 6C
IPI00930073
60.0
14
24.6
82.3
6406 SORD
Sorbitol dehydrogenase
IPI00216057
38.3
11
29.7
110.0
0.35
6410 ANXA1
Annexin A1
IPI00218918
38.7
9
32.9
78.2
0.04
6411 SORD
Sorbitol dehydrogenase
IPI00216057
38.3
10
26.6
92.4
0.39
Protein name: name of each protein in human IPI database. Protein accession number: IPI database accession number. MW: theoretical
molecular weight of identified proteins in kDa. Peptides: number of peptides matched to candidate protein. Sequence coverage: percentage of
identified sequence in relation to complete sequence of known protein. Regulation factor: densitometric ratio between spots in patient
samples compared to respective spots in the control gels. Significance level for regulation was higher than 95% for all spots.
0.05