. 2023 Oct 12;8(10):e012247. doi: 10.1136/bmjgh-2023-012247

Table 2.

Summary of quantitative study methods and results assessing the immunogenicity and cellular immune responses of MAPs

Author, Year	Intervention and sample size (n)	Comparison and sample size (n)	Methods	Immunogenicity findings
Influenza antigen delivered by MAP
Depelsenaire et al., 2021⁴¹	Influenza A/Sing coated MAP (n=5)	Placebo uncoated MAP (n=5)	SII Flow cytometry Immunohistochemistry	All participants receiving influenza MAP were seropositive by day 22, with 80% seroconverted between days 1 and 22. Placebo MAP recipients showed no immune responses. GMTs were similar to the parent study,¹¹ peaking at day 22 (mean=139, 95% CI: 45 to 428).
Fernando et al., 2018¹²	Coated MAP with A/Cali (H1N1)-like antigen on forearm (n=15) Coated MAP with A/Cali (H1N1)-like antigen on deltoid site (n=15)	Fluvax by IM^d injection (n=15) Placebo MAP to forearm (n=5) Placebo MAP to deltoid site (n=5) Saline by IM injection (n=5)	SII HAI assays MN assays Visual analogue scale (0=no pain, 10=worst pain)	Increases in HAI assay GMT were observed on day 7 (p=0.002) and day 21 (p<0.001) in groups receiving the vaccine. GMT was 335 (95% CI: 189 to 593) for influenza MAP to forearm, 160 (95% CI: 74 to 345) for the influenza MAP to deltoid and 221 (95% CI: 129 to 380) for influenza IM injection. By day 21, 80% of influenza MAP to deltoid and IM injection were seroconverted. Sixty per cent of the influenza MAP to forearm group seroconverted by day 21. Seroprotection by day 21 was 100% for the MAP forearm, 87% for the MAP deltoid and 93% for IM injection. MN assays were also used for GMT on day 21, which were 6703 (95% CI: 3534 to 12,713) for MAP forearm, 2211 (95% CI: 1057 to 4624) and 4032 (95% CI: 1854 to 8767). GMTs appeared generally higher in MAPs than IM injection but were insignificant.
Forster et al., 2020¹¹	Part A 15 µg influenza MAP to forearm (n=15) Part B 2.5 µg influenza MAP to forearm (n=20) 5 µg influenza MAP to forearm (n=20) 10 µg influenza MAP to forearm (n=20) 15 µg influenza MAP to forearm (n=20) 15 µg influenza MAP to deltoid (n=20)	Part A Uncoated MAP to forearm (n=15) 15 µg A/Sing by IM injection (n=15) QIV by IM injection (n=15, 1 withdrawal) Part B Uncoated MAP to forearm (n=20) QIV (A/Sing) by IM injection (n=20, 1 withdrawal)	SII HAI assays MN assays ELISA Flow cytometry	No immune responses were observed in participants receiving placebos. There was no difference between low dose (2.5 µg) MAPs and 15 µg IM injections throughout the study. GMT for 2.5 µg MAP was 144.2 (95% CI: 77.9 to 267.0) compared with 15 µg influenza IM injection of 139.3 (95% CI: 79.3 to 244.5, p=0.93). HAI GMTs remained significantly higher during the study between the 10 µg MAP forearm group (309.1, 95% CI: 199.1 to 479.9, p=0.01), 15 µg MAP deltoid group (278.6, 95% CI: 152.7 to 508.1, p=0.03) and IM group (109.3, 95% CI: 59.4 to 200.9) at day 61. GMT fold increases remained significantly higher from baseline in the 15 µg deltoid MAP group (11.3, 95% CI: 6.8 to 18.8) than the IM group (8.9, 95% CI: 5.1 to 15.4, p=0.03). MN GMT was similar to HAI GMT with 10 µg MAP forearm group (18 458, 95% CI: 11 359 to 29 992, p=0.001), 15 µg MAP forearm group (11 362, 95% CI: 6492 to 19 884, p=0.01) and 15 µg MAP deltoid group (13 219, 95% CI: 7096 to 24 626, p=0.02) significantly higher than IM group (3880, 95% CI: 1924 to 7824) at day 22. The 2.5 µg MAP forearm group produced a similar GMT (5301, 95% CI: 2509 to 11 196) to IM group (3880, 95% CI: 1924 to 7824). There were no significant differences in the immune cell responses between influenza MAP and IM vaccine groups.
Hirobe et al., 2015¹⁴	Dissolving MAP with trivalent influenza (n=7)	Trivalent influenza by SC injection (n=20)	Microscopy ELISpot assays HAI assays Haemagglutination assays Sandwich ELISA	H1N1 and H3N2 strains were similar between groups, except B strain were higher in MAP than SC injection. By day 42, seroconversion was higher in the MAP groups compared with SC injection for H1N1 (71% vs 65%) and B strains (43% vs 35%). H3N2 strains had higher seroconversion in MAP than SC injection (43% vs 60%). The same pattern was observed for seroprotection (H1N1: 100% vs 95%, H3N2: 71% vs 95% and B: 86% vs 40%). There were no observed differences in IgA antibodies produced from MAP and SC groups. IFN-γ producing cells increased in both groups, although were slightly higher in the SC group. The MAP group produced HA-specific IFN-γ producing cells and activated T cells, suggesting protection from the influenza virus.
Rouphael et al., 2017¹³	Dissolving MAP with inactivated influenza vaccine self-administered (n=25) Dissolving MAP with inactivated influenza vaccine administered by HCP (n=25)	Inactivated influenza vaccine by IM injection (n=25) Placebo dissolvable MAP (n=25)	HAI assays Food and Drug Administration toxicity grading Surveys	HAI GMTs were similar in both MAP administered by HCP and IM groups at day 28 among all strains: H1N1 (mean=1197, 95% CI: 855 to 1675 compared with mean=997, 95% CI: 703 to 1415, p=0.5), H3N2 (mean=287, 95% CI: 192 to 430 compared with mean=223, 95% CI: 160 to 312, p=0.4) and B strain (mean=126, 95% CI: 86 to 184 compared with mean=94, 95% CI: 73 to 122, p=0.06). By day 28, all groups showed significantly higher seroprotection and seroconversion rates from baseline (p<0.01) but were similar. There was strong evidence that HCP and self-administered MAP groups (65%, 95% CI: 60% to 75%) had a higher seroconversion rate for B strain than IM injection (32%, 95% CI: 15% to 54%, p=0.01). Seroprotection (titre=≥1.4) following vaccination in HCP administered MAP (83%–100%), self-administered MAP (75%–100%) and IM injection (80%–100%) by day 180.
Rouphael et al., 2021⁴²	Dissolving MAP with inactivated influenza vaccine administered by healthcare professional (n=11)	Inactivated influenza vaccine by IM injection (n=11)	HAI assay Neuraminidase inhibition assay Serum antibodies binding assay Innate and T-cell phenotyping assays Influenza virus-specific CD4+ T cells by intracellular cytokine assay Memory B cell analysis by ELISpot assay	HAI GMTs were similar between 1–6 months after vaccination among both groups. GMTs were similar between the three influenza strains (H1N1, H3N2 and B). GMT fold increases were only significantly higher for the B strain (p=0.009). There were no differences between the group rates for seroprotection by day 28 (p=1.00). HAI responses were similar to those observed in the parent study.¹³ Neuraminidase assay GMT showed that strain N1 was higher in MAP at baseline than IM (p=0.04). At day 28, GMT was higher in MAP for strains N1 (p=0.002), N2 (p=0.003) and B (p<0.001) compared with IM. This was also observed on day 2 (N1: p=0.05, B: p=0.02) except for the N2 strain (p=0.11). MAP and IM injection had increased binding affinities at day 28 compared with baseline (MAP: p=0.002, IM: p=0.01). IP-10 increase in the MAP compared with IM (p=0.01) was observed on days 2–3. However, TNF-α was higher in the IM group than in MAP (p=0.04). On day 8, cytokines IL-8 (p=0.02), IL-5 (p=0.034), IL-13 (p=0.013) and MIP-1b (p=0.016) were higher in the MAP group. Following adjustment by Bonferroni correction, these findings were of no significance. Furthermore, there was no difference in the immune cell responses of CD4+ T cell cytokines. At day 28, circulating T follicular helper cells (CD4+, CXCR5+, CXCR3+, ICOS+ and PD-1+) were a higher percentage in the MAP group compared with the IM group (p=0.04).
Japanese encephalitis antigen delivered by MAP
Iwata et al., 2022⁷	Dissolvable MAP containing JEV (high dose) (n=13) Dissolvable MAP containing JEV (low dose) (n=13)	JEV by SC injection (n=13)	Microscopy ELISA Neutralising antibody assays Visual analogue scale (0=no pain, 100=worst pain)	By day 42, participants in all groups (n=39) reached a seroconversion over 1.3 (log10). Seroconversion of ≥1.3 (log10) was observed by day 14 in all high dose MAP participants (n=13), lasting until day 42, with 85% (n=11) on day 189. Over half (n=7) of the SC group seroconverted to ≥1.3 (log10) by day 14. Three-quarters of the low dose MAP group (69%, n=9) and half of the SC group (53.8%, n=7) seroconverted at day 21 and all of these groups seroconverted by day 42. By day 189, neutralising antibodies were observed in 31% (n=4) of the low dose MAP group and 46% (n=6) of SC group. Over days 14, 21 and 189, seroconversion was higher in MAPs than SC injection. Mean titres (log10) were higher in high dose MAPs than in SC injection across the three-time points. At day 42, mean titres in the high dose MAP were 2.55 (SD=0.36), low dose MAP was 2.04 (SD=0.53) and SC injection was 2.08 (SD=0.47). These decreased on day 189 (high dose MAP=1.60, SD=0.39, low dose MAP=0.95, SD=0.32, SC=1.14, SD=0.44). There were significant differences (p=0.0036) between high dose MAP, low dose MAP and SC injection using a contrast coefficient of (−1 to –1, −2). There was no evidence of a correlation between immunogen delivered and titre in the high dose MAP group. There was some evidence of a positive correlation in the low dose MAP group at day 21 (p=0.050) and day 189 (p=0.023).

A/Cali, A/California/07/2009 H1N1; A/Sing, A/Singapore/GP1908/2015 H1N1; ELISpot, enzyme-linked immune absorbent spot; GMT, geometric mean titre; HAI, haemagglutination inhibition; HD-MAP, high-definition microarray patch; IM, intramuscular; JEV, Japanese encephalitis vaccine; MAP, microarray patch; MN, microneutralisation; SC, subcutaneous; SII, Skin irritation index.