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Comparison of the Humoral Immune Response following Both Bacterial Challenge and RNAi of Major Factors on Proliferation of Bartonella quintana in the Human Louse

Human body lice, Pediculus humanus humanus, and head lice, Pediculus humanus capitis, have been hematophagous ectoparasites of humans for thousands of years. Despite being ecotypes, only body lice are known to transmit bacterial diseases to humans, and it appears that lower humoral and cellular immune responses allow body lice to possess a higher vector competence. We previously observed that the transcription level of the defensin 1 gene was up-regulated only in head lice following oral challenge of Bartonella quintana, a causative agent of trench fever, and also that body lice excreted more viable B. quintana in their feces. In this study, we first investigated this differential immune response by performing RNAi to knockdown defensin 1 by dsRNA injection. B. quintana was orally infected 72 h after injection and proliferation was compared at 2 hours (day 0) and day 4 post-infection. At day 0, bacterial cell numbers increased 1.5-fold in defensin 1 (Def1(-)) knocked down head lice compared with non-knocked down, pQE30-dsRNA injected, head lice control. At day 4, Def1(-) knocked down head lice had 2.55-fold more bacterial cells than control head lice and 1.65-fold greater than body lice, indicating that defensin 1 was active in reducing B. quintana cell number in non-knocked down head lice. Second, the levels of cytotoxic reactive oxygen species (ROS) generated by the epithelial cells of the alimentary tract were measured using two general indictors of ROS in both body and head lice at day 1 and day 4 following B. quintana challenge. Challenged body lice showed a 42% and 34% increase in ROS, whereas head lice showed a 70% and 22% increase at day 1 using CM-H2DCFDA and HPF as general indicators, respectively. On day 4, all challenged lice showed similar ROS levels except for body lice which maintained their ROS levels (40% increase using CM-H2DCFDA). Head lice are likely to have multiple immune and/or non-immune factors that suppress B. quintana proliferation, and the production of sustained ROS levels and/or the single knockdown of Defensin 1 is not enough to increase B. quintana proliferation in head lice to that seen in body lice.

Identiferoai:union.ndltd.org:UMASS/oai:scholarworks.umass.edu:masters_theses_2-2284
Date28 October 2022
CreatorsZina, Jake
PublisherScholarWorks@UMass Amherst
Source SetsUniversity of Massachusetts, Amherst
Detected LanguageEnglish
Typetext
Formatapplication/pdf
SourceMasters Theses

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