Nanoparticles (PLGA and Chitosan)-Entrapped ADP-Ribosylation Factor 1 of Haemonchus contortus Enhances the Immune Responses in ICR Mice.

Qiang Qiang Wang,Shakeel Ahmed Lakho,Lixin Xu,Muhammad Waqqas Hasan,Muhammad Ehsan,Javaid Ali Gadahi,Muhammad Haseeb,Ruofeng Yan,Xinchao Liu,Xiangrui Li,Muhammad Ali-Ul-Husnain Naqvi,Xiaokai Song

doi:10.3390/vaccines8040726

Abstract

ADP-ribosylation factor 1 (HcARF1) is one of the Haemonchus contortus (H. contortus) excretory/secretory proteins involved in modulating the immune response of goat peripheral blood mononuclear cells (PBMC). Here, we evaluated the immunogenic potential of recombinant HcARF1 (rHcARF1) against H. contortus infection in Institute of Cancer Research (ICR) mice. Briefly, rHcARF1 was entrapped in poly (D, L-lactide-co-glycolide) (PLGA) and chitosan (CS) nanoparticles (NP) and injected into mice as a vaccine. Fifty-six ICR mice were assigned randomly into seven groups, with eight animals in each group, and they were vaccinated subcutaneously. At the end of the experiment (14th day), the blood and the spleen were collected from euthanized mice to detect lymphocyte proliferation, cytokine analysis, and the production of antigen-specific antibodies. Scanning electron microscope was used to determine the size, morphology, and zeta potential of nanoparticles. Flow cytometry was performed, which presented the increase percentages of CD4+ T cells (CD3e+CD4+), CD8+ T cells (CD3e+CD8+) and dendritic cells (CD11c+CD83+, CD11c+CD86+) in mice vaccinated with rHcARF1+PLGA NP. Immunoassay analysis show raised humoral (Immunoglobulin (Ig)G1, IgG2a, IgM) and cell-mediated immune response (Interleukin (IL)-4, IL-12, and IL-17, and Interferon (IFN)-γ) induced by rHcARF1+PLGA NP. Experimental groups that were treated with the antigen-loaded NP yield higher lymphocyte proliferation than the control groups. Based on these results, we could propose that the rHcARF1 encapsulated in NP could stimulate a strong immune response in mice rather than administering alone against the infection of H. contortus.

Highlights

H. contortus is one of the gastrointestinal nematodes that infect the sheep and goat through feces and cause the disease termed as “haemonchosis” [1]
It revealed that the size of recombinant HcARF1 (rHcARF1)+PLGA NP was 100 ± 10 nm (Figure 1C)
The results demonstrated that IgG1 levels in experimental groups were significantly higher as compared to the phosphate-buffered saline (PBS) group and higher than those in other control groups

Summary

Introduction

H. contortus is one of the gastrointestinal nematodes that infect the sheep and goat through feces and cause the disease termed as “haemonchosis” [1]. The infestation of H. contortus affects thousands of sheep and goats annually, and substantial economic losses to farmers are reported [2]. This parasitic nematode goes into the stomach of the host animal from the herbage while grazing. ARF1 is part of the Ras-related small GTPases family. They are involved in the regulation of vesicular trafficking [6]. It is an essential regulator of the biological process induced by epidermal growth factor [7,8,9]. The replication of the plant RNA virus needed the participation of Arf1 [15]

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