Response In Cervical Lymph Nodes Research Articles

Intranasal Administration of IL-27 Ameliorates Nasal Allergic Responses and Symptoms

Background: Interleukin 27 (IL-27) is an initiator of the Th1 response and inhibits inflammatory responses. In a mouse model of asthma, administration of IL-27 reduced eosinophil numbers in bronchoalveolar lavage fluid and airway hyperresponsiveness. However, it is unclear whether administration of IL-27 can inhibit symptoms of allergic diseases and allergic rhinitis as a therapeutic agent. Therefore, we investigated the in vivo effect of IL-27 on nasal symptoms and allergic rhinitis. Methods: Mice sensitized and challenged with ovalbumin (OVA) antigen received intranasal administration of IL-27. Results: Intranasal administration of IL-27 significantly suppressed the number of sneezes and nasal rubbing movements, the number of eosinophils, OVA-specific T-cell responses in cervical lymph nodes, production of IL-4 and IL-5, and OVA-specific IgE in sera, compared with the administration of PBS alone. The production of IL-10 and IL-35, the percentage of CD25+Foxp3+ cells, and the gene expression of Foxp3 in mice that received intranasal administration of IL-27 were also significantly higher than those in mice that received only PBS. Conclusions: This study showed, for the first time, that intranasal administration of IL-27 inhibited nasal allergic responses and symptoms even after the establishment of allergic rhinitis and suggested that IL-27 is useful as an intranasal therapeutic agent.

Sublingual targeting of STING with 3′3′-cGAMP promotes systemic and mucosal immunity against anthrax toxins

Anthrax is caused by Bacillus anthracis, a zoonotic bacterial pathogen affecting humans and livestock worldwide. The current human anthrax vaccine, anthrax vaccine adsorbed (AVA), is an injected vaccine with a cumbersome administration schedule and fails to promote mucosal immunity. Bacterial enterotoxins, which stimulate production of the cyclic nucleotide cAMP are effective experimental mucosal vaccine adjuvants, but their inherent toxicity has precluded their use in humans. We investigated whether cyclic dinucleotides that target Stimulator of Interferon Gamma Genes (STING) in mammalian cells could represent an alternative to bacterial enterotoxins as adjuvant for sublingual immunization and promotion of mucosal immunity and secretory IgA responses in addition to systemic immunity. We found that sublingual immunization of mice with Bacillus anthracis protective antigen (PA) and the STING ligand 3′3′-cGAMP promotes PA-specific serum IgG Ab responses of the same magnitude as those induced after immunization with PA and the experimental adjuvant cholera toxin (CT). Interestingly, this STING ligand also promoted serum anti-PA IgA and IgA-producing cells in the bone marrow. Furthermore, the saliva of mice immunized with the STING ligand exhibited similar levels of PA-specific IgA Abs as groups immunized with CT as adjuvant. The adjuvant activity of 3′3′-cGAMP was associated with mixed Th1, Th2, and Th17 responses. This STING ligand also induced rapid IFN-β and IL-10 responses in sublingual tissues and cervical lymph nodes, and TGF-β responses in the cervical lymph nodes, which could contribute to promoting IgA responses after sublingual immunization.

Immune response in cervical lymph nodes from patients with primary oral squamous cell carcinoma

Deficient immune response in the cervical lymph nodes of patients with head and neck squamous cell carcinoma may contribute to dissemination of metastatic neoplastic cells. This study evaluates the immune response in cervical lymph nodes from patients with primary oral cavity squamous cell carcinoma (OCSCC). The density of immature (CD1a(+)) and mature (CD83(+)) dendritic cells (DCs), cytotoxic T lymphocytes CD8(+) /perforin(+) (CTLs), and Foxp3(+) regulatory T (Tregs) cells in the lymph nodes of patients with OCSCC without cervical lymph node metastases (LN1) (negative) (n = 10) were identified through immunohistochemistry. From patients with cervical lymph node metastases, samples were obtained of lymph nodes both with (LM2) (positive) (n = 10) and without (LN2) (negative) (n = 10) metastases. The results demonstrated that the number of CD1a(+) and Foxp3(+) cells was significantly higher in the LM2 group than in either the LN1 or the LN2 group. In addition, the number of CD8(+) /perforin(+) and CD83(+) cells was significantly lower in the LM2 group than in the other groups. The results of this study demonstrate a higher density of immature DCs and Tregs cells and a lower density of mature DCs and activated CTLs in metastatic than in non-metastatic lymph nodes. These findings might indicate an immunosuppressive microenvironment, which could be involved in the spread of neoplastic cells to cervical lymph nodes.

Oral macrophage-like cells play a key role in tolerance induction following sublingual immunotherapy of asthmatic mice

AbstractSublingual allergen-specific immunotherapy (SLIT) is a safe and efficacious treatment for type 1 respiratory allergies. Herein, we investigated the key subset(s) of antigen-presenting cells (APCs) involved in antigen/allergen capture and tolerance induction during SLIT. Following sublingual administration, fluorochrome-labeled ovalbumin (OVA) is predominantly captured by oral CD11b+CD11c− cells that migrate to cervical lymph nodes (CLNs) and present the antigen to naive CD4+ T cells. Conditional depletion with diphtheria toxin of CD11b+, but not CD11c+ cells, in oral tissues impairs CD4+ T-cell priming in CLNs. In mice with established asthma to OVA, specific targeting of the antigen to oral CD11b+ cells using the adenylate cyclase vector system reduces airway hyperresponsiveness (AHR), eosinophil recruitment in bronchoalveolar lavages (BALs), and specific Th2 responses in CLNs and lungs. Oral CD11b+CD11c− cells resemble tolerogenic macrophages found in the lamina propria (LP) of the small intestine in that they express the mannose receptor CD206, as well as class-2 retinaldehyde dehydrogenase (RALDH2), and they support the differentiation of interferon-γ/interleukin-10 (IFNγ/IL-10)-producing Foxp3+ CD4+ regulatory T cells. Thus, among the various APC subsets present in oral tissues of mice, macrophage-like cells play a key role in tolerance induction following SLIT.

Toll‐like receptor 2 agonist Pam3CSK4 enhances the induction of antigen‐specific tolerance via the sublingual route

Sublingual immunotherapy (SLIT) has been established in humans as a safe and efficacious treatment for type I respiratory allergies. In this study, we compared three Toll-like receptor (TLR) 2 ligands (Pam3CSK4, Porphyromonas gingivalis lipopolysaccharide and lipoteichoic acid) as potential adjuvants for sublingual allergy vaccines. These molecules were tested in co-cultures of adjuvant-pre-treated dendritic cells (DCs) with murine naïve CD4(+) T lymphocytes. Patterns of cytokine production, phenotype, proliferation and gene expression were analysed by ELISA, cytofluorometry and quantitative PCR, respectively. TLR2 ligands were subsequently tested in a model of SLIT in BALB/c mice sensitized with ovalbumin (OVA). Among the three TLR2 ligands tested, the synthetic lipopeptide Pam3CSK4 is the most potent inducer of IL-12p35 and IL-10 gene expression in murine bone marrow-derived DCs, as well as in purified oral myeloid DCs. Only Pam3CSK4-treated DCs induce IFN-gamma and IL-10 secretion by naïve CD4(+) T cells. Sublingual administration of Pam3CSK4 together with the antigen in BALB/c mice sensitized to OVA decreases airway hyperresponsiveness as well as OVA-specific T-helper type 2 (Th2) responses in cervical lymph nodes dramatically. Pam3CSK4 induces Th1/regulatory T cell responses, and as such, is a valid candidate adjuvant for sublingual allergy vaccines.

Combination of Nasal Lymphotactin and Macrophage Chemoattractant Protein-1 Expressing Plasmids Elicits Antigen-Specific Salivary S-IgA Antibody Immunity

It has been shown that mucosal application of chemokines with protein antigen (Ag) successfully induces mucosal and systemic antibody (Ab) responses, as well as CD4+ T cell responses. We examined whether plasmids expressing lymphotaction (LTN) and/or macrophage chemoattractant proteine-1 (MCP-1) cDNA (pLTN and pMCP-1) exhibit nasal adjuvanticity when administered with protein Ag. When either pLTN or pMCP-1 was employed as a nasal adjuvant in mice, OVA-specific IgG Ab responses were seen in plasma ; however, both failed to induce OVA-specific secretory IgA (S-IgA) Abs in external secretions except anti-OVA IgG and IgA Abs in nasal washes of mice given pLTN. However, mice nasally immunized with a combination of pLTN and pMCP-1 showed OVA-specific S-IgA Ab responses in both saliva and fecal extracts. Further, that combination of chemokines used as a nasal adjuvant induced the highest levels of CD4+ T cell proliferative and Th2-type cytokine responses in cervical lymph nodes (CLN), as compared with a single chemokine nasal administration. In addition, a significantly increased number of CD11c+ dendritic cells (DCs) which expressed co-stimulatory molecules such as MHC II, CD40 and CD86 noted in the nasopharyngeal-associated lymphoreticular tissue (NALT) of mice given pLTN and pMCP-1. These results show that a combination of pLTN and pMCP-1 given as a nasal adjuvant induces optimal mucosal S-IgA Ab responses in submandibular glands, which are mediated by increased number of DCs in the NALT and Th2-type CD4+ T cells in the CLN.

Th2-biased immune response in cervical lymph nodes: Animal models of Graves’ disease might administer autoantigen to body regions where lymphatics drain to cervical lymph nodes

It is believed that cervical lymph nodes have an important impact on Th2-biased immune response to central nervous system (CNS)-derived antigens. Obstruction of these nodes prior to a CNS-infusion of antigen markedly lowers the level of serum antibody response to the antigen. Graves' disease is a Th2-biased autoimmunity in which autoantibodies against thyrotropin receptor stimulate thyroid hormone production. The reasons for the distinct Th2-biased immune response in this hyperthyroidism are not fully understood, whereas most of the other human organ-specific autoimmune diseases are believed to be Th1 mediated. It is suggested that the drainage of the thyroid lymphatics to cervical lymph nodes may play a role in the Th2-polarized immune response of Graves' disease. Thus, the generation of Graves' disease models by immunization may become more feasible if animals are administered the autoantigen to body regions whose lymph is drained to cervical lymph nodes.

TH Cells Primed During Influenza Virus Infection Provide Help for Qualitatively Distinct Antibody Responses to Subsequent Immunization

The quality of the primary Ab-forming cell (AFC) response in cervical lymph nodes and mediastinal lymph nodes of mice to intranasal influenza virus was strongly influenced by viral replicative capacity. IgA secretors were prominent in the early AFC response to infectious virus in mediastinal lymph nodes, while IgG expression was more frequent among isotypically switched AFC in cervical lymph nodes of the same mice; this pattern was reversed in the response to inactivated virus. Influenza viruses A/Puerto Rico/8/34 (A/PR8) and A/X-31 share six of eight genome segments, differing only in hemagglutinin (H1 in A/PR8, H3 in A/X-31) and neuraminidase (N1 in A/PR8, N2 in A/X-31) genes. These viruses therefore elicit extensively cross-reactive TH populations, though their glycoproteins are serologically unrelated. Mice recovered from an A/X-31 infection thus mount a primary B cell response against A/PR8 glycoproteins, when challenged with the latter virus, though this response can call upon memory TH cells. To assess the impact of memory TH populations on a primary Ab response, we compared the AFC response to inactivated A/PR8 in naive mice and mice that had cleared an A/X-31 infection. A/X-31 immune mice mounted a more vigorous AFC response against A/PR8 H1 and N1 glycoproteins than naive animals, when immunized intranasally with inactivated A/PR8. However the distribution of isotypes among H1/N1-specific AFC in lymph nodes of A/X-31-primed mice resembled that of naive mice. Evidently, in this functional context, memory TH cells retained the ability to help Ab responses different in quality from that generated during their primary reaction.