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Vaginal delivery of the recombinant HIV-1 clade-C trimeric gp140 envelope protein CN54gp140 within novel rheologically structured vehicles elicits specific immune responses.

Curran RM, Donnelly L, Morrow RJ, Fraser C, Andrews G, Cranage M, Malcolm RK, Shattock RJ, Woolfson AD - Vaccine (2009)

Bottom Line: CN54gp140 was uniformly distributed within the RSVs and continuously released in vitro in an antigenically intact form over 24h.Vaginal administration to rabbits induced specific serum IgG, and IgG and IgA in genital tract secretions.The RSVs are a viable delivery modality for vaginal immunization.

View Article: PubMed Central - PubMed

Affiliation: The School of Pharmacy, The Queen's University of Belfast, Belfast, BT9 7BL, Northern Ireland, UK. rhonda.curran@qub.ac.uk

ABSTRACT
Rheologically structured vehicle (RSV) gels were developed as delivery systems for vaginal mucosal vaccination with an HIV-1 envelope glycoprotein (CN54gp140). RSVs comprised a mucoadhesive matrix-forming and vaginal fluid absorbing polymer. The mucoadhesive and rheological properties of the RSVs were evaluated in vitro, and the distribution, antigenicity and release of CN54gp140 were analysed by ELISA. CN54gp140 was uniformly distributed within the RSVs and continuously released in vitro in an antigenically intact form over 24h. Vaginal administration to rabbits induced specific serum IgG, and IgG and IgA in genital tract secretions. The RSVs are a viable delivery modality for vaginal immunization.

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Calibration curves for the indirect heterogeneous sandwich ELISA of free-CN54gp140 (□ CN54gp140-spiked 3% RSV, ■ CN54gp140 in PBST, n = 4).
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fig1: Calibration curves for the indirect heterogeneous sandwich ELISA of free-CN54gp140 (□ CN54gp140-spiked 3% RSV, ■ CN54gp140 in PBST, n = 4).

Mentions: No difference was observed between Format 1 and 2 for the construction of CN54gp140-spiked 3% RSV calibration curves, therefore format 2 was selected to ensure minimal use of CN54gp140. A linear calibration plot for CN54gp140 in PBST was observed over the range 0.003–0.05 μg/ml (R2 0.9988) and for the CN54gp140-spiked 3% RSV curve over the range 0.01–0.7 μg/ml (R2 0.9968) (Fig. 1). The LOD of CN54gp140 in PBST was 0.003 μg/ml and the RSD over the entire linear range of the assay of was <8.1%. The LOD of CN54gp140-spiked 3% RSV was 0.01 μg/ml and the RSD was <66% until 0.04 μg/ml (4LOD) and <11% thereafter. The A450 values of the linear range of CN54gp140 in PBST assay were on average 92% higher than those of the CN54gp140-spiked 3% RSV assay. When required samples were diluted to within the linear range of the assay. A CN54gp140-spiked 3% RSV curve was applied to the uniformity of mixing study and stability study and a CN54gp140 in PBST curve was applied to the release studies.


Vaginal delivery of the recombinant HIV-1 clade-C trimeric gp140 envelope protein CN54gp140 within novel rheologically structured vehicles elicits specific immune responses.

Curran RM, Donnelly L, Morrow RJ, Fraser C, Andrews G, Cranage M, Malcolm RK, Shattock RJ, Woolfson AD - Vaccine (2009)

Calibration curves for the indirect heterogeneous sandwich ELISA of free-CN54gp140 (□ CN54gp140-spiked 3% RSV, ■ CN54gp140 in PBST, n = 4).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2824087&req=5

fig1: Calibration curves for the indirect heterogeneous sandwich ELISA of free-CN54gp140 (□ CN54gp140-spiked 3% RSV, ■ CN54gp140 in PBST, n = 4).
Mentions: No difference was observed between Format 1 and 2 for the construction of CN54gp140-spiked 3% RSV calibration curves, therefore format 2 was selected to ensure minimal use of CN54gp140. A linear calibration plot for CN54gp140 in PBST was observed over the range 0.003–0.05 μg/ml (R2 0.9988) and for the CN54gp140-spiked 3% RSV curve over the range 0.01–0.7 μg/ml (R2 0.9968) (Fig. 1). The LOD of CN54gp140 in PBST was 0.003 μg/ml and the RSD over the entire linear range of the assay of was <8.1%. The LOD of CN54gp140-spiked 3% RSV was 0.01 μg/ml and the RSD was <66% until 0.04 μg/ml (4LOD) and <11% thereafter. The A450 values of the linear range of CN54gp140 in PBST assay were on average 92% higher than those of the CN54gp140-spiked 3% RSV assay. When required samples were diluted to within the linear range of the assay. A CN54gp140-spiked 3% RSV curve was applied to the uniformity of mixing study and stability study and a CN54gp140 in PBST curve was applied to the release studies.

Bottom Line: CN54gp140 was uniformly distributed within the RSVs and continuously released in vitro in an antigenically intact form over 24h.Vaginal administration to rabbits induced specific serum IgG, and IgG and IgA in genital tract secretions.The RSVs are a viable delivery modality for vaginal immunization.

View Article: PubMed Central - PubMed

Affiliation: The School of Pharmacy, The Queen's University of Belfast, Belfast, BT9 7BL, Northern Ireland, UK. rhonda.curran@qub.ac.uk

ABSTRACT
Rheologically structured vehicle (RSV) gels were developed as delivery systems for vaginal mucosal vaccination with an HIV-1 envelope glycoprotein (CN54gp140). RSVs comprised a mucoadhesive matrix-forming and vaginal fluid absorbing polymer. The mucoadhesive and rheological properties of the RSVs were evaluated in vitro, and the distribution, antigenicity and release of CN54gp140 were analysed by ELISA. CN54gp140 was uniformly distributed within the RSVs and continuously released in vitro in an antigenically intact form over 24h. Vaginal administration to rabbits induced specific serum IgG, and IgG and IgA in genital tract secretions. The RSVs are a viable delivery modality for vaginal immunization.

Show MeSH