Lung mucosal immunity to NTHi vaccine antigens: Antibodies in sputum of chronic obstructive pulmonary disease patients

Figures & data

Table 1. Patient characteristics.

	Participants N = 40
Age at enrollment in years, mean ± SD	66.3 ± 9.3
Female sex, n (%)	20 (50%)
COPD severity, n (%)
moderate	28 (70%)
severe	12 (30%)
Number of exacerbations during the 12 study months, mean ± SD and median (IQR)	2.9 ± 2.3 and 2 (1–3.5)
Number of exacerbations 1 month post-dose 2 in moderate COPD patients (N = 28), mean ± SD
moderate exacerbations	2.6 ± 2.1
severe exacerbations	2 ± NA
Number of exacerbations 1 month post-dose 2 in severe COPD patients (N = 12), mean ± SD
moderate exacerbations	1.2 ± 0.4
severe exacerbations	0

IQR, interquartile range; N, number; SD, standard deviation.

Table 2. Respiratory function values.

	Moderate patients (N = 28)*		Severe patients (N = 12)
	Range	Mean (SD)	Range	Mean (SD)
FEF25–75	0.37–1.41	0.67 (0.25)	0.20–0.68	0.38 (0.15)
FEV1	0.96–2.51	1.75 (0.44)	0.75–1.57	1.06 (0.26)
FEV1/FVC ratio	42.10–66.70	54.51 (7.24)	31.00–48.90	38.92 (5.88)
FEV1pp	52.30–79.80	64.37 (9.18)	32.30–45.70	37.53 (4.44)
FEV1REV	−208.00–163.00	33.89 (82.01)	−44.00–297.00	66.75 (93.72)
FEV1REV%	−9.06–10.04	2.35 (4.72)	−4.09–27.97	7.51 (9.35)
FVC	1.52–5.16	3.26 (0.88)	2.11–4.13	2.74 (0.60)
FVCpp	64.20–111.50	89.49 (12.60)	58.00–95.10	73.64 (12.27)
PEF	3.00–7.89	5.28 (1.49)	2.21–4.50	3.45 (0.71)
PEFpp	45.50–99.70	76.37 (15.93)	27.0–71.60	49.47 (10.80)

*data from 27 patients for FEV1REV and FEV1REVP.

FEF25–75, forced expiratory flow at 25–75% of forced vital capacity; FEV1, forced expiratory volume in one second; FEV1pp, percentage of predicted value of FEV1; FEV1REV, FEV1 after a reversibility test; FEV1REV%, percent change in FEV1 after a reversibility test; FVC, forced vital capacity; FVCpp, FVC percent predicted; N, number; PEF, peak expiratory flow; PEFpp, percent predicted.

Figure 1. IgG response pre-vaccination vs post-vaccination.

Signals (presented in log10) from IgG antibodies specific for a) PD, b) PE, and c) PilA, normalized for the signals from IgG antibodies against OVA. Signals are from sputum samples collected before the first vaccination (PRE, n = 37) and 30 days after the second vaccination (DAY90, n = 40) with the NTHi vaccine. Each circle represents one sample, measured at dilution 1:10. The horizontal line represents the geometric mean. A value of ≤ 1 indicates background signal (as the specific signal is equal to the signal of the negative control).

Gmean, geometric mean; IgG, immunoglobulin G; NTHi, non-typeable Haemophilus influenzae; OVA, ovalbumin; PD, Protein D; PE, Protein E; PilA, type IV pilin subunit protein.

Figure 2. IgA response pre-vaccination vs post-vaccination.

Signals (presented in log10) from IgA antibodies specific for a) PD, b) PE, and c) PilA, normalized for the signals from IgA antibodies against OVA. Signals are from sputum samples collected before the first vaccination (PRE, n = 37) and 30 days after the second vaccination (DAY90, n = 40) with the NTHi vaccine. Each circle represents one sample, measured at dilution 1:50. The horizontal line represents the geometric mean. A value of ≤1 indicates background signal (as the specific signal is equal to the signal of the negative control).

Gmean, geometric mean; IgA, immunoglobulin A; NTHi, non-typeable Haemophilus influenzae; OVA, ovalbumin; PD, Protein D; PE, Protein E; PilA, type IV pilin subunit protein.

Figure 3. Correlation between IgG in serum and IgG in sputum.

Signals (presented in log10) from DAY90 sputum normalized for the OVA signal (y-axis) and DAY90 serum (on x-axis) of IgG antibodies specific for a) PD, b) PE, and c) PilA. The r is the correlation coefficient between the signals in log10 scale. Each circle represents one individual.

IgG, immunoglobulin G; OVA, ovalbumin; PD, Protein D; PE, Protein E; PilA, type IV pilin subunit protein; r, correlation coefficient.

Figure 4. Correlation of disease severity with antigen-specific antibodies and sputum/serum ratio.

a-c) Signals (presented in log10) from IgG antibodies specific for a) PD, b) PE, and c) PilA, normalized for the signal from IgG antibodies against OVA. Signals are from sputum samples of 12 moderate and 12 severe COPD patients collected 30 days after the second vaccination (DAY90) with the NTHi vaccine. Each circle represents one sample measured at dilution 1:10. The horizontal line represents the geometric mean.

d-f) Ratio of sputum/serum IgG antibodies specific for d) PD, e) PE, and f) PilA. Ratios are from sputum samples of 12 moderate and 12 severe COPD patients collected 30 days after the second vaccination (DAY90) with the NTHi vaccine. Each circle represents the ratio of one sample, measured at dilution 1:10. The horizontal line represents the geometric mean.

COPD, chronic obstructive pulmonary disease; IgG, immunoglobulin G; NTHi, non-typeable Haemophilus influenzae; OVA, ovalbumin; PD, Protein D; PE, Protein E; PilA, type IV pilin subunit protein.

Figure 5. Transudation was detected in sputum from severe COPD patients.

Albumin was measured in sputum of 12 moderate and 12 severe COPD patients as marker for transudation. Each circle represents one sample and is the average of two measurements.

The horizontal line represents the mean. COPD, chronic obstructive pulmonary disease; ND, not detectable.

Data availability statement

GSK makes available anonymized individual participant data and associated documents from interventional clinical studies which evaluate medicines, upon approval of proposals submitted to www.clinicalstudydatarequest.com. To access data for other types of GSK sponsored research, for study documents without patient-level data and for clinical studies not listed, please submit an enquiry via the website. Clinical trial registration is NCT02075541.