PMID- 35179411 OWN - NLM STAT- MEDLINE DCOM- 20220602 LR - 20221207 IS - 1460-2202 (Electronic) IS - 0271-3683 (Linking) VI - 47 IP - 5 DP - 2022 May TI - Elevated Levels of Interleukins, Leukocyte Protein and Cathelicidin Antimicrobial Peptide are Strongly Associated with Early to Mid-Stage of Pythium insidiosum Infection in Rabbit Corneas. PG - 677-687 LID - 10.1080/02713683.2021.2023192 [doi] AB - PURPOSE: Corneal infection in humans caused by Pythium insidiosum can lead to blindness and the host ocular immune response to it is less studied. Herein, we investigate the expression of mediators of innate and adaptive immune responses in a rabbit model. METHODS: P. insidiosum zoospores were injected intracorneally in right eye of the nine New Zealand White rabbits while left eye was injected with 1XPBS. RT-qPCR and multiplex ELISA (mELISA) were used to study the expression of antimicrobial peptides (AMPs) and immune mediators in infected cornea on 3rd, 7th and 9th day of post-infection(PI). STRING-11.0 analysis was used to predict the interactions of immune mediators. mRNA expressions of pathogen recognition receptors (PRRs) were determined in human corneal epithelial cells (HCECs) stimulated with P. insidiosum zoospores. Data was analyzed using one-way ANOVA with Post-Hoc Tukey HSD test and p-value <0.05 was considered significant. RESULTS: mRNA expression assay for IL-1beta, IL-6, IL-8 and Cathelicidin antimicrobial peptide (CAP)-18 showed significant upregulation (p-value < 0.05) on 7th day post-infection (PI) compared to 3rd and 9th day while Leukocyte protein (LeukoP) was elevated significantly on 3rd day followed by 7th and 9th day PI . Only IL-17A among other adaptive immune cytokines showed significant upregulation on 7th day compared to 9th day PI. Expressed in pg/mL, mELISA showed significant higher levels (p-value < 0.05) of IL-1beta, IL-8 in infected tissue in each of the time points compared to control. STRING analysis revealed co-expression of IL-1beta, IL-8 and IL-6. Among PRRs, Dectin 1 and TLR4 showed significant upregulation in HCECs at 12 hrs compared to 6 hrs. CONCLUSION: In the rabbit P. insidiosum keratitis model, innate immune mediators: IL-1beta, IL-8, IL-6, AMPs: LeukoP and CAP-18 are strongly associated in early to mid-stage of corneal infection. Dectin 1 and TLR4 were observed to be associated with recognition of P. insidiosum. FAU - Ahirwar, Lalit Kishore AU - Ahirwar LK AUID- ORCID: 0000-0002-5795-9592 AD - Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, L. V. Prasad Eye Institute, Kallam Anji Reddy Campus, L. V. Prasad Marg, Hyderabad, India. FAU - Sharma, Savitri AU - Sharma S AD - Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, L. V. Prasad Eye Institute, Kallam Anji Reddy Campus, L. V. Prasad Marg, Hyderabad, India. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220322 PL - England TA - Curr Eye Res JT - Current eye research JID - 8104312 RN - 0 (Antimicrobial Cationic Peptides) RN - 0 (Antimicrobial Peptides) RN - 0 (Interleukin-6) RN - 0 (Interleukin-8) RN - 0 (RNA, Messenger) RN - 0 (Toll-Like Receptor 4) RN - 0 (Cathelicidins) SB - IM MH - Animals MH - Antimicrobial Cationic Peptides MH - Antimicrobial Peptides MH - Cornea/metabolism MH - Interleukin-6/metabolism MH - Interleukin-8/genetics/metabolism MH - *Keratitis/metabolism MH - Leukocytes MH - *Pythiosis MH - RNA, Messenger/genetics/metabolism MH - Rabbits MH - Toll-Like Receptor 4/metabolism MH - Cathelicidins OTO - NOTNLM OT - CAP-18 OT - LeukoP OT - Pythium insidiosum OT - cornea OT - innate immune mediators OT - keratitis OT - rabbit EDAT- 2022/02/19 06:00 MHDA- 2022/06/03 06:00 CRDT- 2022/02/18 12:12 PHST- 2022/02/19 06:00 [pubmed] PHST- 2022/06/03 06:00 [medline] PHST- 2022/02/18 12:12 [entrez] AID - 10.1080/02713683.2021.2023192 [doi] PST - ppublish SO - Curr Eye Res. 2022 May;47(5):677-687. doi: 10.1080/02713683.2021.2023192. Epub 2022 Mar 22.