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Insights into the Dynamics of Memory...

Spencer, David A.

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Insights into the Dynamics of Memory, Effector, and Apoptotic Cytotoxic T Lymphocytes in Channel Catfish, Ictalurus punctatus.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Insights into the Dynamics of Memory, Effector, and Apoptotic Cytotoxic T Lymphocytes in Channel Catfish, Ictalurus punctatus./
Author:	Spencer, David A.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2018,
Description:	164 p.
Notes:	Source: Dissertations Abstracts International, Volume: 80-10, Section: B.
Contained By:	Dissertations Abstracts International80-10B.
Subject:	Cellular biology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13870799
ISBN:	9781392025000

Insights into the Dynamics of Memory, Effector, and Apoptotic Cytotoxic T Lymphocytes in Channel Catfish, Ictalurus punctatus.
Spencer, David A.

Insights into the Dynamics of Memory, Effector, and Apoptotic Cytotoxic T Lymphocytes in Channel Catfish, Ictalurus punctatus. - Ann Arbor : ProQuest Dissertations & Theses, 2018 - 164 p.

Source: Dissertations Abstracts International, Volume: 80-10, Section: B.

Thesis (Ph.D.)--The University of Mississippi Medical Center, 2018.

This item must not be added to any third party search indexes.

Cytotoxic T lymphocytes (CTLs) are well studied in mammals, however less is known about the dynamics of memory CTL expansion and exhaustion in teleost fish. This project provides insights into teleost CTL behavior during long-term stimulation using the model alloantigen-specific catfish clonal CTL cell line TS32.15. This cell line was cloned from peripheral blood leukocytes (PBL) isolated from an outbred fish hyper-immunized with catfish allogenic 3B11 B cells, and TS32.15 requires weekly stimulation with irradiated 3B11 B for continuous growth. TS32.15 CTLs are not synchronized, and cultures consist of small cells (10 μm) and large (~30 μm) granular cells. In this study, since some small CTLs bound serum IgM on their surface, and the percentage of IgM binding cells decreased after stimulation, it was initially hypothesized that these CTLs expressed an inhibitory FcjR that was downregulated after stimulation. Instead, here we show that in response to stimulation, small IgM- TS32.15 cells did not immediately lyse targets, but replicated and some differentiated to the large granular morphology typical of mammalian and fish CTLs. Large granular TS32.15 CTLs did not replicate but efficiently lysed 51Cr-labeled 3B11 targets. After lysing targets, or with a prolonged absence of alloantigen stimulation, effector cells transitioned to a small non-cytolytic stage and became apoptotic. Early apoptotic TS32.15 CTLs then became surface positive for IgM. TNP-specific catfish IgM did not bind apoptotic TS32.15 CTLs, and treating apoptotic cells with non-specific lipase reduced IgM binding, indicating natural IgM in serum bind phospholipids exposed on apoptotic CTLs via their Fab region. As might be expected, IgM + TS32.15 CTLs were preferentially cleared by myelocyte-enriched pronephros leukocytes. Transcriptome analysis of small IgM- memory-like TS32.15 CTLs transitioning to the effector stage, large effector TS32.15 CTLs, and IgM+ apoptotic TS32.15 CTLs identified several potential surface markers for further characterization, including fibroblast growth factor receptor-3 (FGFR-3) as a putative marker for activated memory-like TS32.15 CTLs. Additionally, this study found that the IL-10 and IL-17 receptors (IL-10R and IL-17R) were downregulated on large effector TS32.15 CTLs, providing preliminary evidence that IL-10 and IL-17 modulate cell mediated immunity in catfish. Overall, results from this project indicate that in catfish, CTLs undergo clonal expansion and differentiation in response to specific antigen similar to their mammalian counterparts, establish natural IgM as a marker for apoptotic CTLs, and identify putative surface and regulatory proteins for more targeted characterization.

ISBN: 9781392025000Subjects--Topical Terms:

3172791
Cellular biology.

Insights into the Dynamics of Memory, Effector, and Apoptotic Cytotoxic T Lymphocytes in Channel Catfish, Ictalurus punctatus.
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245 1 0 $a Insights into the Dynamics of Memory, Effector, and Apoptotic Cytotoxic T Lymphocytes in Channel Catfish, Ictalurus punctatus.
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300 $a 164 p.
500 $a Source: Dissertations Abstracts International, Volume: 80-10, Section: B.
500 $a Publisher info.: Dissertation/Thesis.
500 $a Advisor: Bengten, Eva.
502 $a Thesis (Ph.D.)--The University of Mississippi Medical Center, 2018.
506 $a This item must not be added to any third party search indexes.
506 $a This item must not be sold to any third party vendors.
520 $a Cytotoxic T lymphocytes (CTLs) are well studied in mammals, however less is known about the dynamics of memory CTL expansion and exhaustion in teleost fish. This project provides insights into teleost CTL behavior during long-term stimulation using the model alloantigen-specific catfish clonal CTL cell line TS32.15. This cell line was cloned from peripheral blood leukocytes (PBL) isolated from an outbred fish hyper-immunized with catfish allogenic 3B11 B cells, and TS32.15 requires weekly stimulation with irradiated 3B11 B for continuous growth. TS32.15 CTLs are not synchronized, and cultures consist of small cells (10 μm) and large (~30 μm) granular cells. In this study, since some small CTLs bound serum IgM on their surface, and the percentage of IgM binding cells decreased after stimulation, it was initially hypothesized that these CTLs expressed an inhibitory FcjR that was downregulated after stimulation. Instead, here we show that in response to stimulation, small IgM- TS32.15 cells did not immediately lyse targets, but replicated and some differentiated to the large granular morphology typical of mammalian and fish CTLs. Large granular TS32.15 CTLs did not replicate but efficiently lysed 51Cr-labeled 3B11 targets. After lysing targets, or with a prolonged absence of alloantigen stimulation, effector cells transitioned to a small non-cytolytic stage and became apoptotic. Early apoptotic TS32.15 CTLs then became surface positive for IgM. TNP-specific catfish IgM did not bind apoptotic TS32.15 CTLs, and treating apoptotic cells with non-specific lipase reduced IgM binding, indicating natural IgM in serum bind phospholipids exposed on apoptotic CTLs via their Fab region. As might be expected, IgM + TS32.15 CTLs were preferentially cleared by myelocyte-enriched pronephros leukocytes. Transcriptome analysis of small IgM- memory-like TS32.15 CTLs transitioning to the effector stage, large effector TS32.15 CTLs, and IgM+ apoptotic TS32.15 CTLs identified several potential surface markers for further characterization, including fibroblast growth factor receptor-3 (FGFR-3) as a putative marker for activated memory-like TS32.15 CTLs. Additionally, this study found that the IL-10 and IL-17 receptors (IL-10R and IL-17R) were downregulated on large effector TS32.15 CTLs, providing preliminary evidence that IL-10 and IL-17 modulate cell mediated immunity in catfish. Overall, results from this project indicate that in catfish, CTLs undergo clonal expansion and differentiation in response to specific antigen similar to their mammalian counterparts, establish natural IgM as a marker for apoptotic CTLs, and identify putative surface and regulatory proteins for more targeted characterization.
590 $a School code: 0805.
650 4 $a Cellular biology. $3 3172791
650 4 $a Immunology. $3 611031
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710 2 $a The University of Mississippi Medical Center. $3 1017852
773 0 $t Dissertations Abstracts International $g 80-10B.
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856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13870799