Cytological observations of transfer of immune complexes via porcine erythrocytes
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Abstract
ABSTRACT
Although the activation of pathogen phagocytosis via complement system has been studied, erythrocyte-phagocyte interactions in pigs are not clearly understood. Therefore, we sought to investigate the ability of porcine erythrocytes to clear immune complexes by using laser confocal microscopy and flow cytometry to observe the immune adhesion of porcine erythrocytes to fluorescent bacilli and the immune presentation process of transferring fluorescent bacilli to macrophages. Isolated porcine alveolar macrophages (PAMs) had uniform morphology and size, and a survival rate of 97.2%. The phagocytosis rate was 98.8%. After WT E. coli was labeled with FITC, the bacteria showed a bright green fluorescence, and the labeling rate was 92.3%. When laser confocal microscopy was utilized to observe the co-incubation system of porcine erythrocytes, PAM, and fluorescent E. coli, the fluorescence intensity of bacilli decreased with increasing observation time and even disappeared. Flow Cytometry examination showed that the average fluorescence intensity of PAMs co-incubated with porcine erythrocytes adhered to WT-E. coli-FITC, was significantly higher than that of normal PAMs. Furthermore, when porcine erythrocytes adhered to WT E. coli were incubated with PAMs, the surface mean fluorescence intensity of porcine erythrocytes was significantly higher than that of the blank control group. This shows that PAMs can competitively bind to the oposinized E. coli adhered to the surface of porcine erythrocytes, and these oposinized pathogens can enter macrophages by the process of phagocytosis，which promoting the internalization of immune complexes or pathogens. During this process, the physical morphology of porcine erythrocytes was not damaged, but the levels of its main functional protein CR1-like were decreased.
Cite this as
2018. Cytological observations of transfer of immune complexes via porcine erythrocytes. PeerJ Preprints 6:e26881v1 https://doi.org/10.7287/peerj.preprints.26881v1note This preprint is not peer-reviewed. You may wish to reference the subsequent peer-reviewed version of this article.
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This is a submission to PeerJ for review.
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Supplemental Information
This file contains the raw data of the study
Figure 4 file contains the original flow cytometry data of the CR1-like quantity test resultsand Figure 5 contains the original flow cytometry data of the E.coli transferation detection.
Additional Information
Competing Interests
The authors declare that they have no competing interests.
Author Contributions
Wei Yin conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the paper, approved the final draft.
Chun Wang conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the paper, approved the final draft.
Kuohai Fan authored or reviewed drafts of the paper, approved the final draft.
Na Sun authored or reviewed drafts of the paper, approved the final draft.
Yaogui Sun authored or reviewed drafts of the paper, approved the final draft.
Hongquan Li conceived and designed the experiments, contributed reagents/materials/analysis tools, approved the final draft.
Animal Ethics
The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):
All the experimental procedures involving animals were approved by the Animal Science and Veterinary Medicine Ethics Committee of Shanxi Agricultural University, China; they strictly adhered to the international biomedical research guidelines (CIOMS and ICLAS, December 2012).
Data Deposition
The following information was supplied regarding data availability:
Raw data is provided in a supplemental file.
Funding
This research was sponsored by National Natural Science Foundation of China (No.31640082) and Shanxi Province Science Foundation for Youth (No.201601D021110). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
