Immunoglobulin A (IgA) antibodies exist in monomeric, dimeric, and secretory forms. Dimerization of IgA depends on a 15-kD polypeptide termed “joining (J) chain,” which is also part of the binding site for an epithelial glycoprotein called “secretory component (SC),” whether this after apical cleavage on secretory epithelia is ligand bound in secretory IgA (SIgA) or in a free form. Uncleaved membrane SC, also called the “polymeric Ig receptor,” is thus crucial for transcytotic export of SIgA to mucosal surfaces, where it interacts with and modulates commensal bacteria and mediates protective immune responses against exogenous pathogens. To evaluate different forms of IgA, we have produced mouse monoclonal antibodies (MAbs) against human J-chain and free SC. We found that J-chain MAb 9A8 and SC MAb 9H7 identified human dimeric IgA and SIgA in enzyme-linked immunoassay and western blot analysis, as well as functioning in immunohistochemistry to identify cytoplasmic IgA of intestinal lamina propria plasmablasts/plasma cells and crypt epithelium of distal human intestine. Finally, we demonstrated that SC MAb 9H7 cross-reacted with rhesus macaque SIgA. These novel reagents should be of use in the study of the biology of various forms of IgA in humans and SIgA in macaques, as well as in monitoring the production and/or isolation of these forms of IgA.
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References
1.
BrandtzaegP: Secretory IgA: Designed for anti-microbial defense. Front Immunol, 2013; 4:222.
2.
BrandtzaegP, KiyonoH, PabstR, and RussellMW: Terminology: Nomenclature of mucosa-associated lymphoid tissue. Mucosal Immunol, 2008; 1:31–37.
3.
FagarasanS, and HonjoT: Intestinal IgA synthesis: Regulation of front-line body defences. Nat Rev Immunol, 2003; 3:63–72.
4.
FernandezMI, PedronT, TournebizeR, Olivo-MarinJC, SansonettiPJ, and PhaliponA: Anti-inflammatory role for intracellular dimeric immunoglobulin A by neutralization of lipopolysaccharide in epithelial cells. Immunity, 2003; 18:739–749.
5.
MacphersonAJ, and UhrT: Induction of protective IgA by intestinal dendritic cells carrying commensal bacteria. Science, 2004; 303:1662–1665.
6.
PetersonDA, McNultyNP, GurugeJL, and GordonJI: IgA response to symbiotic bacteria as a mediator of gut homeostasis. Cell Host Microbe, 2007; 2:328–339.
7.
SuzukiK, MeekB, DoiY, MuramatsuM, ChibaT, HonjoT, et al.: Aberrant expansion of segmented filamentous bacteria in IgA-deficient gut. Proc Natl Acad Sci U S A, 2004; 101:1981–1986.
8.
Battle-MillerK, EbyCA, LandayAL, CohenMH, ShaBE, and BaumLL: Antibody-dependent cell-mediated cytotoxicity in cervical lavage fluids of human immunodeficiency virus type 1—Infected women. J Infect Dis, 2002; 185:439–447.
9.
BluttSE, and ConnerME: The gastrointestinal frontier: IgA and viruses. Front Immunol, 2013; 4:402.
10.
BluttSE, MillerAD, SalmonSL, MetzgerDW, and ConnerME: IgA is important for clearance and critical for protection from rotavirus infection. Mucosal Immunol, 2012; 5:712–719.
11.
GolbySJ, and SpencerJ: Where do IgA plasma cells in the gut come from?. Gut, 2002; 51:150–151.
12.
RenegarKB, SmallPAJr, BoykinsLG, and WrightPF: Role of IgA versus IgG in the control of influenza viral infection in the murine respiratory tract. J Immunol, 2004; 173:1978–1986.
13.
SlackE, BalmerML, FritzJH, and HapfelmeierS: Functional flexibility of intestinal IgA—Broadening the fine line. Front Immunol, 2012; 3:100.
14.
HaynesBF, GilbertPB, McElrathMJ, Zolla-PaznerS, TomarasGD, AlamSM, et al.: Immune-correlates analysis of an HIV-1 vaccine efficacy trial. N Engl J Med, 2012; 366:1275–1286.
15.
BrandtzaegP: The mucosal B-cell system. In: Mucosal Immunology, 4th ed. MesteckyJ, StroberW, RussellMW, KelsallBL, CheroutreH, and LambrechtBN (eds.), Academic Press/Elsevier, Amsterdam, 2015, Chapter 31, pp. 623–681.
16.
WycoffKL: Secretory IgA antibodies from plants. Curr Pharm Des, 2005; 11:2429–2437.
17.
BrandtzaegP: Mucosal and glandular distribution of immunoglobulin components: Differential localization of free and bound SC in secretory epithelial cells. J Immunol, 1974; 112:1553–1559.
18.
BrandtzaegP, and PrydzH: Direct evidence for an integrated function of J-chain and secretory component in epithelial transport of immunoglobulins. Nature, 1984; 311:71–73.
19.
MostovKE, FriedlanderM, and BlobelG: The receptor for transepithelial transport of IgA and IgM contains multiple immunoglobulin-like domains. Nature, 1984; 308:37–43.
20.
BrandtzaegP: Presence of J-chain in human immunocytes containing various immunoglobulin classes. Nature, 1974; 252: 418–420.
21.
JohansenF-E, PeknaM, NorderhaugIN, HanebergB, HietalaMA, KrajciP, BetsholtzC, and BrandtzaegP: Absence of epithelial immunoglobulin A transport, with increased mucosal leakiness, in polymeric immunoglobulin receptor/secretory component-deficient mice. J Exp Med, 1999; 190:915–921.
22.
QuanCP, BernemanA, PiresR, AvrameasS, and BouvetJP: Natural polyreactive secretory immunoglobulin A autoantibodies as a possible barrier to infection in humans. Infect Immun, 1997; 65:3997–4004.
23.
CorthesyB: Recombinant secretory immunoglobulin A in passive immunotherapy. Curr Pharm Biotechnol, 2003; 4:51–67.
24.
FukuyamaY, KingJD, KataokaK, KobayashiR, GilbertRS, OishiK, et al.: Secretory-IgA antibodies play an important role in the immunity to Streptococcus pneumoniae. J Immunol, 2010; 185:1755–1762.
25.
BrandtzaegP: Induction of secretory immunity and memory at mucosal surfaces. Vaccine, 2007; 25:5467–5484.
26.
AmbroseCS, WuX, JonesT, and MalloryRM: The role of nasal IgA in children vaccinated with live attenuated influenza vaccine. Vaccine, 2012; 30:6794–6801.
27.
BelsheRB, GruberWC, MendelmanPM, MehtaHB, MahmoodK, ReisingerK, et al.: Correlates of immune protection induced by live, attenuated, cold-adapted, trivalent, intranasal influenza virus vaccine. J Infect Dis, 2000; 181:1133–1137.
28.
WeintraubJA, HiltonJF, WhiteJM, HooverCI, WycoffKL, YuL, et al.: Clinical trial of a plant-derived antibody on recolonization of mutans streptococci. Caries Res, 2005; 39:241–250.
29.
MantisNJ, PalaiaJ, HessellAJ, MehtaS, ZhuZ, CorthesyB, et al.: Inhibition of HIV-1 infectivity and epithelial cell transfer by human monoclonal IgG and IgA antibodies carrying the b12 V region. J Immunol, 2007; 179:3144–3152.
30.
OtaT, SuzukiY, NishikawaT, OtsukiT, SugiyamaT, IrieR, et al.: Complete sequencing and characterization of 21,243 full-length human cDNAs. Nat Genet, 2004; 36:40–45.
31.
SunagawaK, OmagariD, NishiyamaM, AsanoM, OkuderaM, SugitaniM, et al.: Distinct functional regions of the human polymeric immunoglobulin receptor. Scand J Immunol, 2013; 78:339–344.
32.
ChintalacharuvuKR, and MorrisonSL: Production of secretory immunoglobulin A by a single mammalian cell. Proc Natl Acad Sci U S A, 1997; 94:6364–6368.
33.
LiaoHX, BonsignoriM, AlamSM, McLellanJS, TomarasGD, MoodyMA, et al.: Vaccine induction of antibodies against a structurally heterogeneous site of immune pressure within HIV-1 envelope protein variable regions 1 and 2. Immunity, 2013; 38:176–186.
34.
de Koning-WardTF, FidockDA, ThathyV, MenardR, van SpaendonkRM, WatersAP, et al.: The selectable marker human dihydrofolate reductase enables sequential genetic manipulation of the Plasmodium berghei genome. Mol Biochem Parasitol, 2000; 106:199–212.
35.
NgSK: Generation of high-expressing cells by methotrexate amplification of destabilized dihydrofolate reductase selection marker. Methods Mol Biol, 2012; 801:161–172.
36.
StrausbergRL, FeingoldEA, GrouseLH, DergeJG, KlausnerRD, CollinsFS, et al.: Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. Proc Natl Acad Sci U S A, 2002; 99:16899–16903.
37.
BonsignoriM, HwangKK, ChenX, TsaoCY, MorrisL, GrayE, et al.: Analysis of a clonal lineage of HIV-1 envelope V2/V3 conformational epitope-specific broadly neutralizing antibodies and their inferred unmutated common ancestors. J Virol, 2011; 85:9998–10009.
38.
WuX, ZhouT, ZhuJ, ZhangB, GeorgievI, WangC, et al.: Focused evolution of HIV-1 neutralizing antibodies revealed by structures and deep sequencing. Science, 2011; 333:1593–1602.
39.
SantraS, TomarasGD, WarrierR, NicelyNI, LiaoHX, PollaraJ, et al.: Human non-neutralizing HIV-1 envelope monoclonal antibodies limit the number of founder viruses during SHIV mucosal infection in rhesus macaques. PLoS Pathog, 2015; 11:e1005042.
40.
MoodyMA, AmosJ, GurleyTC, DrinkerM, LuneyKD, MarshallD, et al.: Isolation of human monoclonal antibodies from HIV-uninfected subjects immunized with gp120/NefTat/AS01B by dual-color antigen-specific B cell sorting. AIDS Res Hum Retroviruses, 2010; 26(Suppl):26:OA05.
41.
WhittleJR, ZhangR, KhuranaS, KingLR, ManischewitzJ, GoldingH, et al.: Broadly neutralizing human antibody that recognizes the receptor-binding pocket of influenza virus hemagglutinin. Proc Natl Acad Sci U S A, 2011; 108:14216–14221.
42.
MoldtB, Saye-FranciscoK, SchultzN, BurtonDR, and HessellAJ: Simplifying the synthesis of SIgA: Combination of dIgA and rhSC using affinity chromatography. Methods, 2014; 65:127–132.
43.
TeyBT, and Al-RubeaiM: Effect of Bcl-2 overexpression on cell cycle and antibody productivity in chemostat cultures of myeloma NS0 cells. J Biosci Bioeng, 2005; 100:303–310.
44.
TeyBT, SinghRP, PireddaL, PiacentiniM, and Al-RubeaiM: Influence of bcl-2 on cell death during the cultivation of a Chinese hamster ovary cell line expressing a chimeric antibody. Biotechnol Bioeng, 2000; 68:31–43.
45.
ScearceRM, and EisenbarthGS: Production of monoclonal antibodies reacting with the cytoplasm and surface of differentiated cells. Methods Enzymol, 1983; 103:459–469.
46.
BrandtzaegP: Purification of J-chain after mild reduction of human immunoglobulins. Scand J Immunol, 1975; 4:309–320.
47.
MaB-J, AlamSM, GoEP, LuX, DesaireH, TomarasGM, et al.: Envelope deglycosylation enhances antigenicity of HIV-1 gp41 epitopes for both broad neutralizing antibodies and their unmutated ancestor antibodies. PLoS Pathog, 2011; 7:e1002200.
48.
LiaoHX, SutherlandLL, XiaSM, BrockME, ScearceRM, VanleeuwenS, et al.: A group M consensus envelope glycoprotein induces antibodies that neutralize subsets of subtype B and C HIV-1 primary viruses. Virology, 2006; 353:268–282.
49.
JanewayC, TraversP, WalportM, and ShlomckikJ: Immunobiology: Immune System in Health and Disease, 5th ed. Garland Science, New York, 2001.
50.
LiaoHX, LevesqueMC, NagelA, DixonA, ZhangR, WalterE, et al.: High-throughput isolation of immunoglobulin genes from single human B cells and expression as monoclonal antibodies. J Virol Methods, 2009; 158:171–179.
51.
BrandtzaegP: Human secretory immunoglobulins. V. Occurrence of secretory piece in human serum. J Immunol, 1971; 106:318–323.
52.
BrandtzaegP: Mucosal and glandular distribution of immunoglobulin components. Immunohistochemistry with a cold ethanol-fixation technique. Immunology, 1974; 26:1101–1114.
53.
BrandtzaegP and RognumTO: Evaluation of nine different fixatives. 1. Preservation of immunoglobulin isotypes, J chain, and secretory component in human tissues. Pathol Res Pract, 1984; 179:250–266.
54.
FaschingCE, GrossmanT, CorthesyB, PlautAG, WeiserJN, and JanoffEN: Impact of the molecular form of immunoglobulin A on functional activity in defense against Streptococcus pneumoniae. Infect Immun, 2007; 75:1801–1810.
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