- As Ixodes scapularis nymph tick bites are responsible for most LD cases, this study sought to identify nymph tick saliva proteins associated with B. - Results: We identified a combined 747 tick saliva proteins of uninfected and B. - burgdorferi infected ticks. - burgdorferi infected ticks.. - burgdorferi infection modified protein content in tick saliva to promote its survival at the tick feeding site. - There is evidence that tick saliva proteins facilitate transmission of B. - burgdorferi-infected I. - scapularis nymph tick saliva proteins that are suppressed, induced, enhanced, or not affected in response to B. - Borrelia burgdorferi infected I. - burgdorferi infected ticks (SF2A and SF2B).. - We successfully collected sufficient amount of nymph tick saliva by inserting I. - Likewise, we collected and 9 μg of uninfected tick saliva proteins re- spectively. - To identify proteins in tick saliva, 2 μg of total tick saliva protein per run (in triplicate) was analyzed by LC-MS/. - burgdorferi infected ticks (ST2). - suggesting that secretion of these proteins into tick saliva was induced by B. - 1 Borrelia burgdorferi infection of Ixodes scapularis nymphs induces secretion of a high number of tick saliva proteins. - Total numbers of tick saliva proteins identified at each tick feeding time point (unfed, 12h, 24h, 36h, 48h, 60h, 72h, and fully engorged [SD]) were enumerated. - Green, Yellow, and Purple circles = the number of tick saliva proteins respectively identified in uninfected (UI), Borrelia burgdorferi infected (Bbinf), and found in both. - Borrelia burgdorferi infection modifies secretion dynamics of Ixodes scapularis nymphs tick saliva proteins. - Overall the sum total relative abundance of tick saliva proteins in branch A (signal transduction, antimicrobial, TSP of unknown function, and protein synthesis), branch B (cytoskeletal, extracellular matrix, and glycine rich), nuclear regulation in branch E, lipocalin in branch F were suppressed in saliva of B. - In con- trast, tick saliva proteins in 14 functional categories were secreted at high abundance by B. - Borrelia burgdorferi infection modifies relative abundance of Ixodes scapularis nymph tick saliva proteins. - burgdorferi infection on secretion of rabbit proteins in tick saliva. - To provide context, we have commented on probable functional roles of tick saliva proteins tick transmission of B. - only 2 of the 43 PIs that were identified in saliva of adult I. - burgdorferi infected ticks (Fig. - burgdorferi infected ticks were enhanced in B. - burgdorferi infected ticks (S1 Table). - burgdorferi infected tick saliva which suggests that they play a limited role(s) in B. - burgdorferi infected ticks and was present only in unin- fected tick saliva throughout feeding (Fig. - burgdorferi infected I. - scapularis tick saliva proteome [24], majority of proteases in saliva of I. - It is interesting to note that four of the five cyst- eine proteases in this study were present in saliva of B.. - burgdorferi infected ticks only (Figs. - burgdorferi infected tick saliva. - scapularis genome encodes for other short NPC1-like proteins (XP XP XP and XP that were not found in tick saliva.. - AMPs in tick saliva function to suppress bacteria growth at the tick feeding site and clear microbes in blood meal [54]. - It is notable that, of the eight lipocalins that were enhanced in saliva of B. - burgdorferi infected ticks (Figs. - Some of the immune- related proteins that were enhanced in saliva of B. - burgdorferi infection influenced secretion patterns of tick saliva pro- teins to its benefit. - burgdorferi infected tick saliva [133–135]. - burgdorferi infected tick feeding. - burgdorferi infected ticks (SF5H).. - burgdorferi infected ticks (SF5K). - burgdorferi infected ticks (SF5L). - Secretion dynamics of rabbit proteins in saliva of Borrelia burgdorferi infected ticks Host derived pro- teins have been previously identified in tick saliva pro- teomes during tick feeding [24–26]. - Presence of rabbit proteins in tick saliva might be dis- missed as contamination. - burgdorferi transmission occurs around 48 h, tick saliva proteins that are reverted in abundance during feeding by B. - burgdorferi infected tick saliva from 12-72 h of feeding (ST1) when transmission of the spirochete is expected [33, 45]. - burgdorferi infected tick saliva could indicate its potential role(s) in transmission and dissem- ination through a hematogenous route.. - burgdor- feri infected ticks secreted more tick saliva proteins at high abundance than uninfected ticks, we sought to repeatedly infest rabbits to elicit immunity against B. - burgdorferi in- fection specific tick saliva proteins. - 6, we were curious about antibody response to tick saliva proteins of uninfected and B. - burgdorferi infected (red symbols) ticks. - burgdorferi infected). - This study has described tick saliva proteomes of unin- fected and B. - If this is the case, it is plausible that some of the spirochetes might have migrated to the saliv- ary glands impacting secretion dynamics of tick saliva pro- teins. - burgdorferi colonization of the host. - is also notable that secretion of majority of tick saliva pro- teins by B. - Multiple tick saliva proteins that were enhanced in saliva of B. - This might suggest the possibility that some of the tick saliva proteins that were secreted at high abundance in saliva of B. - burgdorferi infected ticks might be associated with tick saliva exosomes.. - 7 Tick saliva proteins eliciting immunity against Ixodes scapularis nymph feeding are apparently highly expressed in Borrelia burgdorferi infected ticks than uninfected. - burgdorferi infected (red) I. - P/immune = pre-immune, UINF-Ab and 51 = rabbit immune sera to tick saliva proteins of uninfected nymphs. - BbINF- Ab and 49 = rabbit immune sera to tick saliva proteins of B. - burgdorferi infected nymphs. - in naturally infected ticks, and this might have affected secretion of some of the tick saliva proteins in this study.. - This treatment might have re- vealed tick saliva proteins in this study that could be present due to stress response. - Although there is a core set of proteins that are secreted into tick saliva regard- less of the host [22], it is potentially possible that some of the tick saliva proteins identified in this study might not be secreted if ticks are fed on different host such as rodents or humans.. - Therefore, there is a high likelihood that majority of tick saliva proteins in this study will be associated with transmission of B.. - burgdorferi infected nymph tick saliva proteomes allowed us to identify three categories of tick saliva proteins: (i) enhanced, (ii) unmodified, or (iii) sup- pressed in response to B. - speculate that tick saliva proteins that were enhanced in saliva of B. - Equally important, tick saliva proteins that were unmodified with B. - Lastly, we think that tick saliva proteins that were suppressed in response to B. - burgdorferi to adapt to the tick environment before proceeding to tick feeding on rabbits and collecting tick saliva (detailed below).. - Collecting Ixodes scapularis nymph tick saliva. - LC-MS/MS analysis of tick saliva proteins. - scapularis nymph tick saliva proteins were identified using LC-MS/MS analysis as previously described . - Approximately 2 μg of total tick saliva proteins (in triplicate) per feeding time point was processed using LC-MS/MS. - Identification of tick saliva proteins. - Graphic visualization of secretion dynamics of Ixodes scapularis tick saliva proteins. - Our goal was to provoke high antibody titers to tick saliva proteins that were secreted by B. - burgdorferi infected ticks (n=70 per rabbit). - ELISA to quantify immunogenic tick saliva protein expression. - burgdorferi infected ticks during second infestation, we sought to in- vestigate antibody titers to tick saliva proteins in rabbits that were infested by uninfected and B. - burgdorferi infected (n=5). - Profile of uninfected and Borrelia burgdorferi infected Ixodes scapularis nymph tick saliva pro- teins during feeding. - Secretion dynamics of all 747 proteins identified in unin- fected and Borrelia burgdorferi infected Ixodes scapularis nymph tick saliva.. - scapularis nymph tick saliva proteins identified in this study were normalized using the z-score statistics and then used to generate heat maps using heat- map2 function in gplots library using R as described in materials and methods. - Secretion dynamics of protein categories identi- fied in uninfected and Borrelia burgdorferi infected Ixodes scapularis nymph tick saliva. - Tick saliva and its role in pathogen transmission. - Identification of 24h Ixodes scapularis immunogenic tick saliva proteins. - A 24-48 h fed Amblyomma americanum tick saliva immuno-proteome. - Ixodes scapularis tick saliva proteins sequentially secreted every 24 h during blood Feeding. - Time-resolved proteomic profile of Amblyomma americanum tick saliva during feeding. - proteins involved in infection of the tick vector, Ixodes scapularis. - Target validation of highly conserved Amblyomma americanum tick saliva serine protease inhibitor 19. - A snapshot of the Ixodes scapularis degradome
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