About RecombiMAb anti-mouse LPAM-1 (Integrin α4β7) The DATK32-CP069 monoclonal antibody is a recombinant, chimeric version of the original DATK32 antibody. The variable domain sequences are identical but the constant region sequences have been switched from rat IgG2a, κ to mouse IgG2a, κ for use in murine models. Species-matched chimeric antibodies exhibit regulated effector functions—including Fc receptor binding and complement activation—and result in less immunogenicity and formation of anti-drug antibodies (ADAs) than xenogenic antibodies in animal models. The highly controlled sequence and lack of genetic drift in recombinant antibodies provide more reliable and reproducible results over hybridoma derived antibodies. The DATK32 monoclonal antibody reacts with mouse LPAM-1 also known as integrin alpha 4 beta 7. The 130 kDa integrin β7 chain associates with the 150 kDa integrin α4 (CD49d) chain to form LPAM-1, a member of the Ig superfamily. LPAM-1 is expressed by peripheral lymphocytes, small subsets of thymocytes, and bone marrow progenitors. LPAM-1 binds VCAM-1 (CD106), MAdCAM-1, and fibronectin and facilitates lymphocyte adhesion and migration to the intestine and associated lymphoid tissues. The DATK32 antibody has been reported to block LPAM-1-mediated cell adhesion in vivo. RecombiMAb anti-mouse LPAM-1 (Integrin α4β7) Specifications IsotypeMouse IgG2a, κ Recommended Isotype Control(s)RecombiMAb mouse IgG2a isotype control, unknown specificity Recommended Dilution BufferInVivoPure pH 7.0 Dilution Buffer ImmunogenTK1 cells Reported Applicationsin vivo Integrin α4β7 neutralization Flow cytometry For information on in vivo applications, please contact technicalservice@bioxcell.com FormulationPBS, pH 7.0 Contains no stabilizers or preservatives Endotoxin<1EU/mg (<0.001EU/μg) Determined by LAL gel clotting assay Aggregation<5% Determined by SEC Purity>95% Determined by SDS-PAGE Sterility0.2 μm filtration ProductionPurified from HEK293 cell supernatant in an animal-free facility PurificationProtein G Molecular Weight150 kDa StorageThe antibody solution should be stored at the stock concentration at 4°C. Do not freeze. Application ReferencesRecombiMAb anti-mouse LPAM-1 (Integrin α4β7) (CLONE: DATK32-CP069)Duc D, Vigne S, Bernier-Latmani J, Yersin Y, Ruiz F, Gaïa N, Leo S, Lazarevic V, Schrenzel J, Petrova TV, Pot C (2019). "Disrupting Myelin-Specific Th17 Cell Gut Homing Confers Protection in an Adoptive Transfer Experimental Autoimmune Encephalomyelitis" Cell Rep 29(2):378-390.e4. PubMedMultiple sclerosis (MS) is a common autoimmune disease of the CNS. Although an association between MS and inflammatory bowel diseases is observed, the link connecting intestinal immune responses and neuroinflammation remains unclear. Here we show that encephalitogenic Th17 cells infiltrate the colonic lamina propria before neurological symptom development in two murine MS models, active and adoptive transfer experimental autoimmune encephalomyelitis (EAE). Specifically targeting Th17 cell intestinal homing by blocking the α4β7-integrin and its ligand MAdCAM-1 pathway impairs T cell migration to the large intestine and dampens EAE severity in the Th17 cell adoptive transfer model. Mechanistically, myelin-specific Th17 cells proliferate in the colon and affect gut microbiota composition. The beneficial effect of blocking the α4β7-integrin and its ligand MAdCAM-1 pathway on EAE is interdependent with gut microbiota. Those results show that disrupting myelin-specific Th17 cell trafficking to the large intestine harnesses neuroinflammation and suggests that the gut environment and microbiota catalyze the encephalitogenic properties of Th17 cells.Bemark M, Hazanov H, Strömberg A, Komban R, Holmqvist J, Köster S, Mattsson J, Sikora P, Mehr R, Lycke NY (2016). "Limited clonal relatedness between gut IgA plasma cells and memory B cells after oral immunization" Nat Commun . PubMedUnderstanding how memory B cells are induced and relate to long-lived plasma cells is important for vaccine development. Immunity to oral vaccines has been considered short-lived because of a poor ability to develop IgA B-cell memory. Here we demonstrate that long-lived mucosal IgA memory is readily achieved by oral but not systemic immunization in mouse models with NP hapten conjugated with cholera toxin and transfer of B1-8(high)/GFP(+) NP-specific B cells. Unexpectedly, memory B cells are poorly related to long-lived plasma cells and less affinity-matured. They are α4β7-integrin(+)CD73(+)PD-L2(+)CD80(+) and at systemic sites mostly IgM(+), while 80% are IgA(+) in Peyer's patches. On reactivation, most memory B cells in Peyer's patches are GL7(-), but expand in germinal centres and acquire higher affinity and more mutations, demonstrating strong clonal selection. CCR9 expression is found only in Peyer's patches and appears critical for gut homing. Thus, gut mucosal memory possesses unique features not seen after systemic immunization.Rosser EC, Oleinika K, Tonon S, Doyle R, Bosma A, Carter NA, Harris KA, Jones SA, Klein N, Mauri C (2014). "Regulatory B cells are induced by gut microbiota-driven interleukin-1β and interleukin-6 production" Nat Med 20(11):1334-9. PubMedRegulatory B cells (Breg cells) differentiate in response to inflammation and subsequently restrain excessive immune responses via the release of interleukin-10 (IL-10). However, the precise inflammatory signals governing their differentiation remain to be elucidated. Here we show that the gut microbiota promotes the differentiation of Breg cells in the spleen as well as in the mesenteric lymph nodes. Perturbation of the gut microbiome imposed either by antibiotic treatment or by changes in the sterility of housing conditions reduces the number and function of Breg cells. Following the induction of arthritis, IL-1β and IL-6 are produced only in conventionally housed mice and both cytokines directly promote Breg cell differentiation and IL-10 production. Mice lacking IL-6 receptor (IL-6R) or IL-1 receptor 1 (IL-1R1) specifically on B cells have a reduced number of IL-10-producing B cells and develop exacerbated arthritis compared to control animals. Thus, in response to inflammatory signals induced by both the gut flora and arthritis, Breg cells increase in number and restrain excessive inflammation.
