title: Fine-Needle Aspiration Biopsy of the Lymph Node: A Novel Tool for the Monitoring of Immune Responses after Skin Antigen Delivery.
abstract: Assessment of immune responses in lymph nodes (LNs) is routine in animals, but rarely done in humans. We have applied minimally invasive ultrasound-guided fine-needle aspiration of the LN to a before-and-after study of the immune response to intradermally delivered Ag in healthy volunteers (n = 25). By comparison with PBMCs from the same individual, LN cells (LNCs) were characterized by reduced numbers of effector memory cells, especially CD8(+) TEMRA cells (3.37 ± 1.93 in LNCs versus 22.53 ± 7.65 in PBMCs; p = 0.01) and a marked increased in CD69 expression (27.67 ± 7.49 versus 3.49 ± 2.62%, LNCs and PBMCs, respectively; p < 0.0001). At baseline, there was a striking absence of IFN-γ ELISPOT responses to recall Ags (purified protein derivative, Tetanus toxoid, or flu/EBV/CMV viral mix) in LN, despite strong responses in the peripheral blood. However, 48 h after tuberculin purified protein derivative administration in the ipsilateral forearm resulting in a positive skin reaction, a clear increase in IFN-γ ELISPOT counts was seen in the draining LN but not in PBMCs. This response was lost by 5 d. These data suggest that the low levels of effector memory cells in the LN may explain the low background of baseline ELISPOT responses in LNs as compared with PBMCs, and the appearance of a response after 48 h is likely to represent migration of effector memory cells from the skin to the LN. Hence, it appears that the combination of intradermal Ag administration and draining LN sampling can be used as a sensitive method to probe the effector memory T cell repertoire in the skin.
authors.affiliation: Diabetes Research Group, Institute for Molecular and Experimental Medicine, Cardiff University School of Medicine, Cardiff CF14 4XN, Wales, United Kingdom; tatovicd@cardiff.ac.uk.
authors.forename: Danijela
authors.initials: D
authors.lastname: Tatovic
authors.affiliation: Radiology Department, University Hospital of Wales, Cardiff CF14 4XN, Wales, United Kingdom; and.
authors.forename: Philippa
authors.initials: P
authors.lastname: Young
authors.affiliation: NanoPass Technologies, Nes Ziona 74036, Israel.
authors.forename: Efrat
authors.initials: E
authors.lastname: Kochba
authors.affiliation: NanoPass Technologies, Nes Ziona 74036, Israel.
authors.forename: Yotam
authors.initials: Y
authors.lastname: Levin
authors.affiliation: Diabetes Research Group, Institute for Molecular and Experimental Medicine, Cardiff University School of Medicine, Cardiff CF14 4XN, Wales, United Kingdom;
authors.forename: F Susan
authors.initials: FS
authors.lastname: Wong
authors.affiliation: Diabetes Research Group, Institute for Molecular and Experimental Medicine, Cardiff University School of Medicine, Cardiff CF14 4XN, Wales, United Kingdom;
authors.forename: Colin M
authors.initials: CM
authors.lastname: Dayan
mesh_terms: D000293:Adolescent; D000328:Adult; D015703:Antigens, CD / Q000276:immunology; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology; D000956:Antigens, Viral / Q000008:administration & dosage; D044963:Biopsy, Fine-Needle / Q000295:instrumentation / Q000379:methods*; D002465:Cell Movement; D058501:Enzyme-Linked Immunospot Assay; D005260:Female; D006801:Humans; D007156:Immunologic Memory; D016130:Immunophenotyping; D007271:Injections, Intradermal; D007371:Interferon-gamma / Q000096:biosynthesis; D037181:Lectins, C-Type / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D008198:Lymph Nodes / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D008213:Lymphocyte Activation; D008297:Male; D008875:Middle Aged; D012867:Skin / Q000187:drug effects / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D013745:Tetanus Toxoid / Q000008:administration & dosage; D014373:Tuberculin / Q000008:administration & dosage; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D000956:Antigens, Viral; D007371:Interferon-gamma; D037181:Lectins, C-Type; D013745:Tetanus Toxoid; D014373:Tuberculin; D000099192:CD69 Antigens