Adverse event
n %
(95% CI)
n %
(95% CI)
n %
(95% CI)
n %
(95% CI)
n %
(95% CI)
n % (95% CI) 21 95.5 11 50.0 6 75.0 2 25.0 5 71.4 0 0.0 At least one
AE (77.2-99.9) (28.2-71.8) (34.9-96.8) (3.2-65.1) (29.0-96.3) (0.0-41.0) Abdom. pain 1 4.5 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Abdom. pain 1 4.5 0 0.0 0 0.0 0 0.0 1 14.3 0 0.0 upper (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.4-57.9) (0.0-41.0) Anorexia 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Anxiety 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Appendicitis 1 4.5 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Arthralgia 1 4.5 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Back pain 2 9.1 0 0.0 1 12.5 0 0.0 0 0.0 0 0.0 (1.1-29.2) (0.0-15.4) (0.3-52.7) (0.0-36.9) (0.0-41.0) (0.0-41.0) Cellulitis 1 4.5 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Cellulitis 1 4.5 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 staphylococcal (0.1-22.8) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Cough 1 4.5 0 0.0 0 0.0 0 0.0 2 28.6 0 0.0
(0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (3.7-71.0) (0.0-41.0) Diarrhoea 1 4.5 1 4.5 1 12.5 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.1-22.8) (0.3-52.7) (0.0-36.9) (0.0-41.0) (0.0-41.0) Decreased 5 22.7 0 0.0 0 0.0 0 0.0 1 14.3 0 0.0 CD4 count (7.8-45.4) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.4-57.9) (0.0-41.0) Dysmenorrhoe
a
1 4.5 1 4.5 1 12.5 0 0.0 0 0.0 0 0.0
(0.1-22.8) (0.1-22.8) (0.3-52.7) (0.0-36.9) (0.0-41.0) (0.0-41.0) Dyspepsia 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Epistaxis 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Fatigue* 2 9.1 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 (1.1-29.2) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Feeling hot 2 9.1 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 (1.1-29.2) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Flatulence 0 0.0 0 0.0 1 12.5 1 12.5 0 0.0 0 0.0 (0.0-15.4) (0.0-15.4) (0.3-52.7) (0.3-52.7) (0.0-41.0) (0.0-41.0) Gastroenteritis 1 4.5 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Genital herpes 0 0.0 0 0.0 1 12.5 0 0.0 0 0.0 0 0.0 (0.0-15.4) (0.0-15.4) (0.3-52.7) (0.0-36.9) (0.0-41.0) (0.0-41.0) Headache* 5 22.7 2 9.1 2 25.0 1 12.5 0 0.0 0 0.0 (7.8-45.4) (1.1-29.2) (3.2-65.1) (0.3-52.7) (0.0-41.0) (0.0-41.0)
simplex
(0.1-22.8) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Hot flush 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Injection site 2 9.1 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 pruritus (1.1-29.2) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Insomnia 1 4.5 0 0.0 0 0.0 0 0.0 1 14.3 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.4-57.9) (0.0-41.0) Musculoskelet
al
1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0
chest pain (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Migraine 3 13.6 2 9.1 0 0.0 0 0.0 0 0.0 0 0.0 (2.9-34.9) (1.1-29.2) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Myalgia* 4 18.2 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (5.2-40.3) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Nasal 0 0.0 0 0.0 1 12.5 0 0.0 0 0.0 0 0.0 congestion (0.0-15.4) (0.0-15.4) (0.3-52.7) (0.0-36.9) (0.0-41.0) (0.0-41.0) Nasopharyngit
is
4 18.2 1 4.5 2 25.0 0 0.0 2 28.6 0 0.0
(5.2-40.3) (0.1-22.8) (3.2-65.1) (0.0-36.9) (3.7-71.0) (0.0-41.0) Nausea 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Neck pain 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0
(0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Night sweats 1 4.5 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) 0.1-22.8 (0.0-36.9) 0.0-36.9 (0.0-41.0) (0.0-41.0) Oral herpes 1 4.5 0 0.0 0 0.0 0 0.0 1 14.3 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.4-57.9) (0.0-41.0) Oropharyngeal 2 9.1 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 pain (1.1-29.2) (0.1-22.8) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Ovarian cyst 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Pain in 2 9.1 0 0.0 0 0.0 0 0.0 1 14.3 0 0.0 extremity (1.1-29.2) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.4-57.9) (0.0-41.0) Palpitations 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Pterygium 0 0.0 0 0.0 0 0.0 0 0.0 1 14.3 0 0.0 (0.0-15.4) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.4-57.9) (0.0-41.0) Pyrexia 2 9.1 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (1.1-29.2) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Rash papular 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Rash pruritic 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Rectal 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 haemorrhage (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Subcutaneous 0 0.0 0 0.0 0 0.0 0 0.0 1 14.3 0 0.0
Tinnitus 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Vertigo 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Visual 1 4.5 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 impairment (0.1-22.8) (0.0-15.4) (0.0-36.9) (0.0-36.9) (0.0-41.0) (0.0-41.0) Vomiting 1 4.5 0 0.0 1 12.5 0 0.0 0 0.0 0 0.0 (0.1-22.8) (0.0-15.4) (0.3-52.7) (0.0-36.9) (0.0-41.0) (0.0-41.0)
At least one AE = at least one adverse event experienced (regardless of the MedDRA Preferred Term); N = number of subjects with at least one administered dose; n/% = number/percentage of subjects reporting at least once the symptom;
95% CI= exact 95% confidence interval; * AEs that started outside of the 7-day follow-up period for solicited AEs were classified as unsolicited AEs.
Group timing N mean s.d. min Q1 median Q3 max M72/AS01 pre (D0) 22 611.2 266.4 243 416 585 651 1427
PI (D30) 22 626.6 279.0 217 465 566 713 1443 PII (D60) 22 599.2 224.7 257 457 567 679 1093 PII (D210) 22 566.8 194.7 263 426 548.5 680 1002 AS01 pre (D0) 8 434.1 138.8 245 318.5 437 561 595 PI (D30) 8 425.6 152.9 274 283.5 410.5 527.5 688 PII (D60) 8 449.8 148.8 324 339 368 572.5 715 PII (D210) 8 459.9 118.2 316 377 433 534 675
Saline pre (D0) 7 878.3 235.6 682 704 811 1050 1336 PI (D30) 7 962.6 549.3 458 750 799 978 2158 PII (D60) 7 923.9 285.1 548 745 877 1062 1453 PII (D210) 7 987.4 499.6 531 741 822 1138 2045
N, number of subjects with available results (the Total Vaccinated Cohort); s.d., standard deviation; Q1/Q3, first/third quartiles; min/max, minimum/maximum; pre (D0), blood sample taken before the first vaccination dose at the first study visit (Day 0); PI (D30), post first vaccination (Day 30); PII (D60), post second vaccination (Day 60);
PII (D210), post second vaccination (Day 210).
gating, then a lymphocyte gating (FSC-A/SSC -A), followed by discrimination of the CD4+ and CD8+ T cells from the “morphological” gating window. (b) CD4 + T cells were analyzed with respect to the production of CD40L, IL-2, TNF- α and IFN- γ. The unstimulated (medium) samples show background levels of cytokine production.
When restimulated with the M72 peptide pool, the production of CD40L, IL-2 and TNF-α by CD4+ T cells and lower production of IFN- γ was detected. The numbers in the quadrant gates of the plots denominate each distinct population based on their cytokine production. Samples from the same subject are shown, with responses at one month post second vaccination. Results shown are representative of the range of responses seen with all subjects studied.
FSC-A
SSC-A
FSC-H CD8
CD4
CD40L IL-2
CD40L IL-2
M ed ium M 72
CD4
CD4 CD4 CD4
CD4
CD4 CD4 CD4
TNF-α TNF-α IFN-γIFN-γ
(a)
(b)
months after the second dose (D60 and D210) for BCG-naive and BCG-vaccinated subjects of the M72/AS01 group (N=6 and N=16, respectively). Data are reported as the percentages of M72-specific CD4+ T cells expressing (after in vitro stimulation) at least two immune markers among IFN-γ, IL-2, TNF-α and CD40L (A), or, at least one immune marker among IFN-γ, IL-2, TNF-α and CD40L (‘total’) or at least one cytokine among IFN-γ, IL-2 and TNF-α
(
B) of all CD4+ T cells, with first and third quartiles, and the minimum/maximum values measured.BCG-naïve
BCG-vaccinated D0 D30
D60
D210 D0 D30 D60
D210 D0 D30 D60
D210 D0 D30 D60
D210
Immune marker
+ M72-specific CD4
+ T cells (%)
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6
Total IL-2+ TNF-α+ IFN-γ+
BCG-naïve BCG-vaccinated A.
B.
Max
Min Q1 Q3 Median
Total IL-2+ TNF-α+ IFN-γ+
D0 D30 D60
D21
0 D0 D30
D60 D21
0 D0 D30
D60 D21
0 D0 D30
D60 D21 0.0 0
0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6
D0 D30 D60
D210 D0 D30
D60 D210
M72-specific CD4
+ T cells expressing at least 2 markers (%)
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4
concentration; CI, confidence interval. Anti-M72 IgG antibody responses are shown for all groups (A) and for the BCG-naive and BCG-vaccinated subjects of the M72/AS01 group (B). The ELISA cut-off was set at 2.8 EU/ml.
A.
assay cut-off
D0 D30 D60
D210 D0 D30 D60
D210 D0 D30 D60
D210
Anti-M72 IgG (GMC EU/ml, with 95% CI)
1 10 100 1000
Saline M72/AS01 AS01
D0 D30 D60
D210 D0 D30
D60 D210 1
10 100 1000
BCG-naïve BCG-vaccinated B.
