Secretory immunoglobulin A sIgA is a promising candidate for enteric therapeutics applications, and several sIgA-based constructs are currently being developed by groups utilizing clarified Chinese ...hamster ovary CHO cell culture supernatants. To the monoclonal antibody downstream processing typically entails chromatography-based purification processes beginning with Protein A chromatography. In this paper, aqueous two-phase systems ATPS were employed for the preliminary purification of secretory immunoglobulin A sIgA monoclonal antibody mAb from clarified CHO-cell culture supernatants. A 24 full factorial design was utilized. The influence of various process parameters such as pH, PEG molecular weight MPEG, PEG concentration CPEG, and phosphate salt concentration CPHO, on the sIgA partition coefficient K sIgA and the recovery index Y in the PEG phase were evaluated. The Elisa assay revealed that, in the ATPS conditions tested, sIgA mAb was mostly detected in PEG upper phase. Run 14 with the highest sIgA activity exhibited the following conditions: MPEG 8.000 g/mol, CPEG 12,5 %, pH 7,0 and CPHO 10 %, and a sIgA K of 94.50 and a recovery index Y of 33.52 %. The proposed platform provides straightforward implementation, yields comparable results, and offers significantly improved economics for manufacturing sIgA mAb biotherapeutics.
Latar belakang. Kolostrum susu sapi atau Bovine colostrum mengandung berbagai growth factor dan immune factor, salah satunya secretory IgA (sIgA) yang dengan jumlah signifikan dapat menghalangi ...adhesi patogen ke membrane mukosa dan menghambat kolonisasi sehingga bermanfaat untuk mengobati penyakit di saluran pencernaan. Penelitian yang telah dilakukan di Indonesia belum banyak membahas secara spesifik hubungan sIgA yang didapatkan dari kolostrum susu sapi sebagai terapi tambahan diare akut pada anak. Tujuan. Mengetahui pengaruh pemberian kolostrum susu sapi terhadap durasi diare akut dehidrasi ringan sedang dan kadar sekretori IgA pada anakMetode. Penelitian eksperimental yang dilaksanakan di Puskesmas dan Rumah Sakit di kota Padang. Penelitian dimulai dari bulan februari tahun 2022 sampai November 2022. Data terkumpul 30 sampel masing-masing pada kelompok kontrol yang mendapatkan terapi standar WHO dan kelompok intervensi yang mendapatkan terapi standar who ditambah kolostrum susu sapi. Dilakukan pengamatan terhadap durasi diare akut serta pemeriksaan terhadap kadar sIgA.Hasil. Terjadi pemendekan durasi diare secara signifikan antara kelompok intervensi dan kelompok kontrol sebesar 11,93 jam (p=0,021). Terdapat perbedaan kadar sIgA yang signifikan antara sebelum dan sesudah pemberian kolostrum susu sapi pada kelompok intervensi (p=0,003).Kesimpulan. Pemberian kolostrum susu sapi dapat memperpendek rerata durasi diare akut dan meningkatkan rerata kadar sIgA secara bermakna. Pemberian kolostrum susu sapi ini dapat disarankan sebagai terapi adjuvan dalam tatalaksana diare akut pada anak.
•Alhagi honey polysaccharides could increase pIgR secretion in Caco-2 cell in vitro.•Alhagi honey polysaccharides could significantly induce DCs maturation in vitro.•Alhagi honey polysaccharides ...could enhance sIgA in gut of cyclophosphamide-mice.•Alhagi honey polysaccharides could activate the DCs in cyclophosphamide-mice.
It remains a huge challenge to recover the intestine immune function for the treatment of intestinal mucosal damage from chemotherapy with cyclophosphamide (CY). Alhagi honey polysaccharide (AH) has immunomodulation pharmacological activity, but the effect and mechanism on the intestinal immune system of CY-mice remain unclear.
In this experiment, the immunomodulatory activity of AH on intestinal immune in CY-mice and its mechanism of regulating the intestinal immune system was investigated.
The experiment studied the immunomodulatory activity of AH on the intestinal immune system and its mechanism for the first time from in vitro and in vivo experiments. We investigated the immunomodulatory effects of AH on Caco-2 and dendritic cells (DCs) in vitro by using western blot (WB), flow cytometry, quantitative real-time PCR (qPCR), and ELISA methods. In vivo experiment, the immunosuppressive mouse model was established through being given intraperitoneal injection with CY (80 mg/kg) for 3 days. Then, mice oral administration of 800 mg/kg AH and 40 mg/kg levamisole hydrochloride for a week. Immunofluorescence, flow cytometry, ELISA, qPCR and WB were applied to examine the immunomodulatory activity of AH on the intestinal immune function of CY-mice, as well as the function of AH on the concentration of SCFAs in cecum by Gas chromatographic analysis.
In vitro experiments, AH could significantly stimulate the expression of pIgR protein in Caco-2. It could also induce the DCs maturation and release the cytokines to regulate the immune response. In vivo experiments, AH could remarkably stimulate the DCs maturation and secrete more CCL20 to recruit DCs, then induce the T (CD4+ and CD8+) and B cells proliferation and activation. Moreover, it could further induce T helper cells to differentiate and secrete cytokines to enhance the secretion of sIgA. Furthermore, it also directly activated DCs and released cytokines to increase the content of pIgR, J-chain, and IgA+ cells in intestine, thereby enhancing the secretion of sIgA to protect the intestine. In addition, AH could obviously strengthen the SCFAs production in cecum to regulate the intestinal immune dysfunction induced by CY.
In summary, oral administrated AH exhibits great benefits for treating CY-induced intestinal immunosuppression, and the mechanism of action mainly involves sIgA, DCs, SCFAs.
Latar belakang. Air susu ibu (ASI) berperan penting dalam sistem kekebalan innate bayi baru lahir. Kolostrum telah diketahui mengandung antibodi terbesar yaitu sIgA yang berasal dari ...Entero-broncho-Mammary Pathway. Sekretori IgA kolostrum merupakan imunomodulator perkembangan sistem imun bayi baru lahir. Antibodi pada ASI sering dihubungkan dengan perlindungan bayi terhadap infeksi sampai usia 6 bulan. Masih perlu diketahui peranan sIgA kolostrum terhadap kejadian infeksi Balita.Tujuan. Membuktikan hubungan antara kadar sIgA kolostrum dengan frekuensi infeksi umum dan infeksi saluran pernapasan akut (ISPA) pertahun pada Balita. Mengetahui faktor lain yang berhubungan dengan infeksi Balita secara umum.Metode. Studi prospektif pada 53 Balita usia 3 tahun dengan riwayat kehamilan cukup bulan, lahir sehat beserta ibunya. Pengumpulan ASI pada hari ke 2-3 postpartum, sIgA diukur dengan teknik ELISA. Data didapat dari kuesioner dengan kunjungan rumah atau pertelepon. Analisis statistik menggunakan uji Mann-Whitney, chi-square dan Fisher.Hasil. Rerata frekuensi infeksi umum kelompok riwayat kadar sIgA kolostrum rendah (<120mg/dL) lebih sering dibanding kelompok kontrol (5,9 vs 4,2, p=0,03), demikian juga frekuensi ISPA lebih sering dibanding kontrol (5,2 vs 3,8, p=0,041). Terdapat hubungan antara kepadatan hunian dengan sering infeksi, p=0,01.Kesimpulan. Balita dengan riwayat sIgA kolostrum ibu rendah (<120mg/dL) lebih sering mengalami infeksi saluran pernapasan akut dan infeksi secara umum dibanding kontrol. Kepadatan hunian berhubungan dengan riwayat sering infeksi.
The level of secretory immunoglobulin A (SIgA) in saliva is associated with the risk of upper respiratory infection and has been identified as a condition indicator for athletes and an indicator of ...mucosal immune function in the elderly. In addition, it has been suggested that non-specific polyreactive SIgA is present in human saliva and colostrum. The purpose of this study was to establish a new detection method of polyreactive salivary SIgA. Using saliva samples from eight healthy individuals, indirect ELISA was used to detect anti-LPS antibodies (LPS ELISA), and competitive ELISA was used to detect LPS antibodies after competition with DNA (LPS-DNA ELISA). The difference in SIgA concentration between LPS ELISA and LPS-DNA ELISA is thought to be the concentration of the polyreactive SIgA, which responds to both DNA and LPS. First, we determined the concentration for coating LPS and DNA as 10 μg/ml, and the saliva sample dilution was determined as 1/2. The concentration of anti-LPS antibody contained in the saliva sample, which is strongly reactive to LPS, was defined as the standard. Compared to the SIgA concentration determined by LPS ELISA, SIgA concentration of LPS-DNA ELISA did not decrease in two samples, but was reduced in 6 samples. The decrease in SIgA concentration is thought to be the concentration of polyreactive SIgA that reacts with both DNA and LPS. Notably, we found sample-to-sample differences in the concentration of polyreactive SIgA.
Enterohemorrhagic
(EHEC) has consistently been one of the foremost foodborne pathogen threats worldwide based on the past 30 years of surveillance. EHEC primarily colonizes the bovine ...gastrointestinal (GI) tract from which it can be transmitted to nearby farm environments and remain viable for months. There is an urgent need for effective and easily implemented pre-harvest interventions to curtail EHEC contamination of the food and water supply. In an effort to address this problem, we isolated single-domain antibodies (V
Hs) specific for intimin, an EHEC adhesin required for colonization, and designed chimeric V
H fusions with secretory IgA functionality intended for passive immunotherapy at the mucosal GI surface. The antibodies were produced in leaves of
with production levels ranging between 1 and 3% of total soluble protein.
assembly of all subunits into a hetero-multimeric complex was verified by co-immunoprecipitation. Analysis of multivalent protection across the most prevalent EHEC strains identified one candidate antibody, V
H10-IgA, that binds O145:Hnm, O111:Hnm, O26:H11, and O157:H7. Fluorometric and microscopic analysis also indicated that V
H10-IgA completely neutralizes the capacity of the latter three strains to adhere to epithelial cells
. This study provides proof of concept that a plant-produced chimeric secretory IgA can confer cross-serotype inhibition of bacterial adhesion to epithelial cells.
Segmented filamentous bacteria (SFB) are commensal organisms that grow by anchoring a specialized holdfast structure to the intestinal walls of a variety of animals. Interaction of SFB with Peyer's ...patches in mice promotes the post-natal maturation of the immune system. We previously reported that the colonization of SFB in humans mainly occurs by 36 months of age, and is difficult to be detected afterward. In this study, we measured the level of SFB in intestinal fluids of human children. SFB were found via qPCR to represent a small fraction of the whole SFB-positive microbiota (10
SFB in 10
total bacteria). Bacteria with filamentous segmented morphology were observed in intestinal fluids via fluorescent
hybridization, and from gut biopsies via scanning electron microscopy. SFB-specific DNA and peptide fragments were also identified via multiple displacement amplification PCR and mass spectrometry. There was an overall positive correlation between the presence of SFB and the titer of total secretory immunoglobulin A (sIgA), which is more apparent in intestinal fluids of the age group of 8-36 months. Afterward there was a decline of SFB in numbers correlated with a reduction of total sIgA. RT-qPCR analysis of the terminal ileal biopsies revealed that the expression of Th17 pathway genes were induced in SFB-positive samples, while the markers of T and B cell receptor signaling pathways were also upregulated. Collectively, these data suggest that SFB is a rare member of microbiota, and may play an important role in the development of human gut immunity.
Summary
Introduction
Among sensitized infants, those with high, as compared with low levels, of salivary secretory IgA (SIgA) are less likely to develop allergic symptoms. Also, early colonization ...with certain gut microbiota, e.g.
Lactobacilli
and
Bifidobacterium
species, might be associated with less allergy development. Although animal and
in vitro
studies emphasize the role of the commensal gut microbiota in the development of the immune system, the influence of the gut microbiota on immune development in infants is unclear.
Objective
To assess whether early colonization with certain gut microbiota species associates with mucosal and systemic immune responses i.e. salivary SIgA and the spontaneous Toll‐like receptor (TLR) 2 and TLR4 mRNA expression and lipopolysaccharide (LPS)‐induced cytokine/chemokine responses in peripheral blood mononuclear cells (PBMCs).
Methods
Fecal samples were collected at 1 week, 1 month and 2 months after birth from 64 Swedish infants, followed prospectively up to 5 years of age. Bacterial DNA was analysed with real‐time PCR using primers binding to
Clostridium difficile
, four species of bifidobacteria, two lactobacilli groups and
Bacteroides fragilis
. Saliva was collected at age 6 and 12 months and at 2 and 5 years and SIgA was measured with ELISA. The PBMCs, collected 12 months after birth, were analysed for TLR2 and TLR4 mRNA expression with real‐time PCR. Further, the PBMCs were stimulated with LPS, and cytokine/chemokine responses were measured with Luminex.
Results
The number of
Bifidobacterium
species in the early fecal samples correlated significantly with the total levels of salivary SIgA at 6 months. Early colonization with
Bifidobacterium
species, lactobacilli groups or
C. difficile
did not influence TLR2 and TLR4 expression in PBMCs. However, PBMCs from infants colonized early with high amounts of
Bacteroides fragilis
expressed lower levels of TLR4 mRNA spontaneously. Furthermore, LPS‐induced production of inflammatory cytokines and chemokines, e.g. IL‐6 and CCL4 (MIP‐1β), was inversely correlated to the relative amounts of
Bacteroides fragilis
in the early fecal samples.
Conclusion
Bifidobacterial diversity may enhance the maturation of the mucosal SIgA system and early intense colonization with
Bacteroides fragilis
might down‐regulate LPS responsiveness in infancy.
The immunocompetence handicap hypothesis (ICHH) contends that testosterone-dependent, masculine traits are honest signals of the bearer's ability to withstand immunosuppression, yet empirical studies ...testing the ICHH have produced mixed results. The present research addressed the relationship between masculine vocal parameters and immunity in two samples: young adult men from the US and Canada (N = 267) and adolescent males from Bolivia (N = 44). All participants provided saliva samples, assayed for a biomarker of mucosal immunity associated with respiratory disease resistance: secretory immunoglobulin-A (sIgA). Participants also contributed speech samples, analyzed for two sexually dimorphic aspects of the human voice: 1) fundamental frequency (fo; perceived as pitch) and vocal tract length (VTL; estimated using formant frequencies). Multiple regression models were used to assess the association between masculine vocal parameters and sIgA. Results indicated that lower fo and longer VTL were associated with higher sIgA levels. That is, both adult and adolescent males with more masculine voices showed better mucosal immunity (and therefore disease resistance) than those with less masculine voices. Although this accords with several previous studies on voice and sIgA, it contrasts with standard ICHH-derived predictions. We suggest that life-history theory provides a better conceptual framework for understanding the association between testosterone-dependent secondary sexual characteristics and immunity as it shifts the focus from immunosuppression to efficient immunoredistribution.
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