Immune Response Targeting Sjögren's Syndrome Antigen Ro52 Suppresses Tear Production in Female Mice.

Marta Trzeciak,Umesh Deshmukh,Harini Bagavant,Joanna Papinska

doi:10.3390/ijms19102935

Marta Trzeciak, Umesh Deshmukh + Show 2 more

Open Access

https://doi.org/10.3390/ijms19102935

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Abstract

Autoantibodies reactive against Ro52 are present in 70% of Sjögren’s syndrome patients and are associated with higher disease severity. However, their role in causing aqueous deficient dry eye, a major cause for morbidity in Sjögren’s syndrome, is unclear. To investigate whether immune responses targeting Ro52 contribute towards the dry eye, male and female NZM2758 mice were immunized with recombinant Ro52. Tear production was measured by the phenol red thread test. Sera were analyzed for anti-Ro52 levels by immunoprecipitation. Lacrimal glands were evaluated for inflammatory foci and IgG deposits. Our results showed that, although all mice generated anti-Ro52 antibodies, only females developed a significant drop in tear production. None of the mice developed severe lacrimal gland inflammation, and female mice with anti-Ro52 showed higher levels of IgG deposits within their glands. Passive transfer of anti-Ro52 sera caused reduced tear production in female mice, but not in males. This study demonstrates for the first time that immune responses initiated by Ro52 induce aqueous dry eye, and this may be driven by anti-Ro52 antibodies. Furthermore, the sexual dimorphism in glandular dysfunction suggests that the lacrimal glands in females are more susceptible to autoantibody-mediated injury.

Highlights

Dry eye is one of the major complications of Sjögren’s syndrome, affecting almost 90% of patients [1]
IInn thhee currentt study, to determine whether immune response to Ro52 affects lacrimal gland function, tear production in immunized mice was measured by using the phenol red thread test (Figure 1)
Female mice immunized with recombinant Ro52 protein showed a signifificant droopp ((4477%%,pp

Summary

Introduction

Dry eye is one of the major complications of Sjögren’s syndrome, affecting almost 90% of patients [1]. Dry eye disease in Sjögren’s syndrome is caused by reduced fluid secretion by the lacrimal gland, as well as by the evaporative loss of tear volume [2]. Multiple factors, such as autoantibodies, inflammatory cell infiltrates, and proinflammatory cytokines can potentially influence lacrimal gland function [3,4,5,6]. Autoimmune mechanisms of dry eye disease have been investigated in rodent models using immunization with crude lacrimal gland extracts [7,8,9], purified kallikrein 1b22 [10], and acinar cell microparticles [11] as antigens. PPrreevviioouuss wwoorrkk ffrroomm oouurr llaabboorraattoorryy hhaass ddeemmoonnssttrraatteedd tthhaatt iimmmmuunnee rreessppoonnsseess aaggaaiinnsstt wwhhoollee RRoo5522 oorr aafrfargagmmenent tcocnotnatianiinnigngitsitcsoicleodil-ecdo-iclodilomdoaminariendurecdeupcielocpairlopcinarep-iinndeu-icnedduscaeldivasatiloivnaitniomn iicne m[25ic,2e6[]2.5T,2h6e].saTlihvearsyalgilvaanrdy dgylasnfudndctyiosfnuinncttihoins eixnptehriismeexnptearlimmoednetal lwmasodaneltiwboadsya-nmteibdoiadtye-dmaenddiawteads adnedpewndasendteopnenthdeenact toivnatthioenaocftiivnantaioteniomfminunnaittey.imTomeuluncitiyd.aTteotehleurcoidleatoef tahnetir-oRloe5o2finandtri-yReoy5e2, iinn tdhriys esytued, yin, twheis ismtumduy,nwizeedimNmZuMni2z7e5d8 NmZicMe2o7f58bomthiceseoxfebsowthitshexaesrewcoitmhbainreacnotmmbainltaonset mbianldtoinsegbpinrodtienign p(MroBtePi)n-m(MouBsPe)-Rmoo5u2 sfeusRioon52pfruosteioinn (pMroBtPei-nm(RMoB52P)-.mCRono5tr2o)l. mCoicnetrwoel rme iicmemwuenreiziemdmwuitnhizMedBPwaitlhonMe.BIPn asolomnee.eIxnpesroimmeenetxsp, earhimigehnttsit,ear hanigtih-Rtiot5e2r aimntmi-Runo5e2seimrummuwneasseprausmsivwelays tpraansssifveerrlyedtrianntosfeNrrZeMd 2in75to NmZalMe 2a7n5d8 fmemalealaenmd ifceem. aOleurmdicaet.aOshuorwdatthaasthaonwtit-hRaot5a2nrtie-dRuoc5e2dretdeaurcepdrotedaurcptiroonduocntliyoninonfelmy ianlefemmiaclee. mAnicteib. oAdnytidbeopdoysidtieopnoswitiitohninwtihtheinlatchreimlaaclrigmlaanldglaapnpdeaaprspetoarsbetorbeesproenspsiobnlesibfoler ftohrethinediuncdtuiocntioonf ogflagnldanudlaurladrydsfyusnfuctnioctnio. nO. uOr urresruesltusltssusguggegstestthtahtatfefmemalaelelalcarcirmimalalgglalannddssaarree mmoorree ssuusscceeppttiibbllee ttoo iimmmmuunnee--mmeeddiiaatteedd iinnjjuurryy

Results

Proteins and High Titer Immune Sera

Histopathology

Passive Transfer of Rabbit Serum into Adjuvant Treated Mice

Statistical Methods