Innate immune cells---- one time migration preprogrammed homing properties
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1 Innate immune cells---- one time migration preprogrammed homing properties neutrophils---acute inflammation monocytes---subacute/chronic inflammation eosinophils---parasitic or allergic inflammation natural killer cells mast cells Adaptive immune cells recirculate and are reprogrammed during antigen responses B and T cells: Naïve lymphocytes---programmed to home and recirculate through secondary lymphoid tissues Memory/effector lymphocytes---reprogrammed for tissue and microenvironment specific trafficking (gut skin joint CNS lymph nodes etc.) Dendritic (professional antigen presenting) cells 1
2 fetal lymphocytes lymphoid tissue homing skin draining homing lymphocytes gut homing adult memory/effector intestinal lymph cells T cells naïve lymphocytes organized lymphoid tissues Antigen specificity influences accumulation but not recruitment gut Peyer s patches spleen lymph nodes inflamed skin circulating memory lymphocytes memory and effector lymphocytes: an army of specialists Husband and Gowans 2
3 specialized post-capillary high endothelial venules (HEV) periph. lymph node Peyer s patch peripheral lymph node Peyer s patch (GALT) 3
4 Mucosal vascular addressin (MAdCAM-1) peripheral lymph node Peyer s patch (GALT) 4
5 a4b7 hi memory cells embody immune memory for gastrointestinal antigens Cutaneous lymphocyte antigen (CLA) 5
6 cutaneous lymphomas express CLA 6
7 Skin-homing (CLA+) and gut-homing (a4b7 hi) T cell populations are mutually exclusive control treatment inflammatory bowel disease a4b7 integrin blockade Ringler et al 7
8 1. neutrophils express L-selectin, and use it in recruitment to inflamed tissues (Lewinsohn, 1987) 2. neutrophils like lymphocytes can use L-selectin to bind to HEV (but do not extravasate via HEV) 3. skin homing lymphocytes use E-selectin in chronic inflammation (Picker, 199), but neutrophils do so in acute inflammation. => additional steps must be involved! CD18 integrins mediate neutrophil sticking (Arfors and Harlan) CD18 integrins mediate neutrophil sticking (Arfors and Harlan) Anti-CD18 QuickTime and a Sorenson Video decompressor are needed to see this picture. QuickTime and a Sorenson Video decompressor are needed to see this picture. Granulocytes accumulate in reperfusion injury Anti-CD18 integrin inhibits sticking but cells still roll 8
9 9
10 receptors responsible for initial contact (tethering) are expressed on microvilli receptors specialized for later steps are excluded from villi Microvillous presentation facilitates interaction under flow α4β7 LFA-1 U. von Andrian 1
11 MAdCAM-CHO X X X X X X X X X X X X X X QuickTime and a Sorenson Video decompressor are needed to see this picture. Anti-alpha 4 MAb in multiple sclerosis Natalizumab for Relapsing Multiple Sclerosis Chaudhuri A., Behan P. O., Miller D. H., O'Connor P. W. N Engl Med 23; 348: , Apr 17, 23 11
12 lymphoid tissue homing How do tissue-specific subsets of effector and memory lymphocytes arise? Daniel. Campbell, Chang Kim skin homing gut homing Technical issues: Low frequency of antigen-specific lymphocytes >OVA-specific TCR system (DO-11.1) Lymphocyte recirculation makes it difficult to definitively determine a given cell s site of activation >examine phenotype at day 1-2, when Ag-specific cells are sequestered >used a4b7 and TECK responsiveness as indication of gut homing properties, P-selectin binding as systemic HR CD4+ T cells activated in cutaneous vs. intestinal lymph nodes differentially express P-lig and α4β7 1 PLN 1 MLN P-lig LPS+OVA α4β7 LPS alone PLN MLN %α4β7 hi MLN PLN % P-lig Cell divisions tissue-specific induction of homing specialization begins within 1-2 cell divisions α4β7 P-lig D. Campbell and C. Kim 12
13 Conclusions Expression of adhesion and chemokine homing receptors is rapidly induced during the primary antigen response in lymphoid tissues The selectivity of homing properties is determined by the microenvironment at the site of antigen stimulation and initial proliferation B and T lymphocytes recirculate naïve lymphocytes---programmed to home and recirculate through secondary lymphoid tissues Homing specificity is reprogrammed during antigen responses memory and effector lymphocytes display specialization of tissue and microenvironment-specific trafficking (gut skin joint CNS lymph nodes etc.)?role of dendritic cells?role for subsequent homeostatic modulation of homing properties in long term memory populations Specificity of lymphocyte-endothelial cell interactions function of the combination of receptors a cell expresses multi-step adhesion cascade Acknowledgements: Part 1 - Tissue-specific lymphocyte subsets: epithelial chemokines in the segregation of mucosal IgA ASC Eric Kunkel, Nicole Lazarus, Eddie Bowman, Ken Youngman Part 2 - Induction of homing specificity during the naïve to memory/effector transition Daniel. Campbell, Chang Kim MLN PLN OVA-specific T cells 2 days after immunization: chemotaxis to IP-1 (general inflammatory chemokine) MLN PLN TECK (intestinal chemokine) Part 3 - Separable populations of helper and DTH CD4+ effector cells Daniel. Campbell (Chang H. Kim) Migration (% of input) tissue-specific induction of chemotactic specificity stimulation in gut but not peripheral lymph nodes induces responsiveness to the intestinal chemokine TECK 13
14 14
15 tissue and microenvironment-selective lymphocyteendothelial cell recognition sequences tissue and microenvironment-selective lymphocyteendothelial cell recognition sequences * * * C * * * * C * Peyer's patch (T) L-selectin/ CCR7/SLC others? Peyer's patch (T) L-selectin/ CCR7/SLC others? Peyer's patch (B) L-selectin/??? CXCR5/BLC? Peyer's patch (B) L-selectin/??? CXCR5/BLC? peripheral LN (T cells) L-selectin/PNAd CCR7/SLC others? peripheral LN (T cells) L-selectin/PNAd CCR7/SLC others? gut lamina propria??? CCR9/TECK gut lamina propria??? CCR9/TECK inflamed skin CLA/E-selectin CCR4/TARC CTACK inflamed skin CLA/E-selectin CCR4/TARC CTACK BALT L-selectin/ α4β1/vcam-1 CCR7/SLC α4β1/vcam-1 others? BALT L-selectin/ α4β1/vcam-1 CCR7/SLC α4β1/vcam-1 others? 15
16 tissue and microenvironment-selective lymphocyteendothelial cell recognition sequences two means of regulating integrin-dependent adhesion * * C * * avidity Peyer's patch (T) L-selectin/ CCR7/SLC others? Peyer's patch (B) L-selectin/??? CXCR5/BLC? clustering peripheral LN (T cells) L-selectin/PNAd CCR7/SLC others? gut lamina propria??? CCR9/TECK resting (no adhesion) affinity inflamed skin BALT CLA/E-selectin L-selectin/ α4β1/vcam-1 CCR4/TARC CCR7/SLC α4β1/vcam-1 CTACK others? conformational changes Mn++ treatment induces high affinity LFA1 PMA induces adhesion through avidity changes A B cell adhesion no Mn 2+ Mn 2+ pretreatment soluble *ICAM binding no Mn 2+ Control PTX anti-lfa-1 Mn++ induces LFA-1-dependent lymphocyte adhesion to ICAM-1 upregulates integrin affinity >soluble ICAM-1 binding C D cell adhesion PMA no agonist Time (min) soluble *ICAM binding PMA no agonist phorbol esters (PKC stimulation) induce LFA-1-dependent lymphocyte adhesion to ICAM-1 no affinity upregulation >no binding of soluble ICAM Mn 2+ -pretreatment Time (min) 16
17 A cell adhesion to surface ICAM chemokines trigger soluble ICAM binding rapid adhesion through affinity regulation no agonist Time (sec) DF-1 α no agonist SDF-1 MIP-3β no agonist MIP3b (ELC) 6-C-kine no agonist SLC B solluble ICAM-1 bound Time (sec.) SDF-1α no agonist SDF-1 MIP-3β no agonist MIP3b (ELC) 6-C-kine no agonist SLC chemokine affinity triggering is mediated by PTXsensitive G protein signaling C 4 no agonist D Control Control SDF-1α MIP-3β 6-C-kine anti-lfa-1 no agonist SDF-1α MIP-3β 6-C-kine PTX soluble ICAM-1 binding is LFA-1 dependent soluble ICAM-1 binding is PTX sensitive Boosting the frequency of antigen-specific CD4+ T cells to detectable levels OVA-specific cells are sequestered in lymphoid tissues until day three after systemic OVA immunization 2 Balb/c % K OVA-specific T cells in blood CD4 DO Days post immunization Transfer OVA-specific TCR transgenic T cells from DO11.1 mice into nontransgenic Balb/c recipients Balb/c after transfer Blood PLN MLN day 2 K1-26 CFSE 17
18 Karl Arfors and ohn Harlan (1987) -anti-cd18 inhibits sticking of neutrophils in inflamed venules => rolling and sticking are mediated by distinct mechanisms von Andrian, utila, Kishimoto, Arfors et al (199/1) -anti-l-selectin inhibits rolling of neutrophils in vivo, and prevents integrin dependent arrest Generalized multistep model: -sequential rolling, activation and arrest could be a general mechanism for leukocyte-endothelial recognition. 18
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