Increased Antibody Production Research Articles

Analysis of antibody production in Saccharomyces cerevisiae: effects of ER protein quality control disruption.

One of the main limitations for heterologous protein production in the yeast Saccharomyces cerevisiae is the protein-folding capacity in the endoplasmic reticulum (ER). Accumulation of unfolded proteins triggers the unfolded protein response (UPR), which resolves the stress by increasing the capacity for protein folding and removal of unfolded proteins by the ER-associated degradation (ERAD) system. In order to analyze the influence of ERAD on production of a human IgG, we disrupted ERAD at different stages by deletion of the HTM1, YOS9, HRD1, HRD3, or UBC7 gene, with or without a disruption of the UPR by deletion of the IRE1 gene. All deletion strains were viable and did not exhibit a growth phenotype under normal growth conditions. Deletion of HTM1 resulted in a small increase in antibody production, whereas a small decrease in antibody production was observed in the Δhrd1, Δhrd3, and Δubc7 yeast strains, and a stronger decrease in the Δyos9 yeast strain. Deletion of the IRE1 gene had contrasting effects in the ERAD mutants, with a strongly decreased production in wild-type cells and partially reversed effects in combination with the Δhtm1 or the Δyos9 deletions. In order to study IgG clearance from the ER, an assay was developed using the inhibitory effect of glucose on the GAL1 promoter that is driving IgG expression. The Δyos9Δire1and Δhtm1Δire1 strains showed a delayed IgG clearance from the cells, showing that removal of components for the generation and recognition of the glycan signal needed for ERAD-mediated protein degradation might increase the IgG ER residence time.

The immunologic effects of estrogen on psoriasis: A comprehensive review.

BackgroundImmunological changes in pregnancy are associated with improvements in some pre-existing immune-mediated skin diseases. Estrogen has been hypothesized to contribute to these changes by creating a shift from Th1 and Th17 to Th2 immunity. As this hypothesis would predict, psoriasis (a primarily Th17 mediated immune disease) tends to improve during pregnancy. However, the precise mechanism by which estrogen induces immunological change in psoriasis remains poorly understood.ObjectiveTo summarize the immunologic effects of estrogen as they relate to psoriasis during pregnancy.MethodsWe performed an English-language PubMed search of articles from September 2004 to September 2014 combining the key terms "psoriasis," "estrogen," "autoimmune disease," and "pregnancy."ResultsEstrogen appears to up-regulate Th2 cytokines and down-regulate Th1 and Th17 cytokines. This shift was initially observed in murine systems, which showed decreased mixed lymphocyte reactions of splenocytes and increased antibody production during pregnancy. Antigen stimulated splenocytes produced fewer Th1 cytokines and more Th2 cytokines in pregnant mice. IL17 producing T cells were significantly decreased in healthy pregnancies compared to non-pregnant controls.LimitationsThis review is limited by the paucity of studies evaluating immunological changes of psoriasis in pregnancy among human subjects.ConclusionsIncreased estrogen production in pregnancy is associated with decreased Th1 and Th17 cytokine production. While estrogen may be responsible for some of these immune shifts resulting in disease improvement, there remains no definitive evidence to prove the hypothesis that estrogen is responsible for such improvement.

Chronic liver fibrosis triggers autoantibody production by B cells in a MyD88-dependent pathway (BA4P.126)

Abstract Our recent finding suggests that B cells contribute to liver fibrosis through hepatic stellate cells (HSCs) activation. While antibodies produced by the activated intrahepatic B cells contribute to autoimmunity is not known, mixed cryoglobulinemia and other B cell disorders are the leading extrahepatic manifestations of advanced liver diseases in humans. Here, our aim was to determine whether increased antibody production during liver fibrosis could potentially lead to systemic autoimmunity by production of autoantibodies and immune complexes (IC). We studied the role of B cells in autoimmunity using a carbon tetrachloride (CCl4)-induced mouse model of fibrosis and a genetically engineered multidrug resistance gene 2 knock out (Mdr2-/-) mice, which constitutively develop progressive liver fibrosis. In both fibrosis models, the frequency of B cells in the liver was elevated and B cells exhibited increased state of activation as measured by CD44 and CD86 expressions, constitutive IgG production and elevated TNF-a production. By using antinuclear antibody (ANA) staining with HEp-2 stabilized substrate and C1q standard ELISA, we detected the presence of antinuclear antibodies (ANA) and IC respectively in the serum of C57BL/6 mice treated with CCl4 for six weeks and Mdr2-/- mice at 25 week. Interestingly, the targeted deletion of MyD88 signaling in B cells during liver fibrosis resulted in reduced ANA in the serum suggesting the critical role of MyD88 in systemic autoimmunity.

Mechanisms of immunity in post-exposure vaccination against Ebola virus infection.

Ebolaviruses can cause severe hemorrhagic fever that is characterized by rapid viral replication, coagulopathy, inflammation, and high lethality rates. Although there is no clinically proven vaccine or treatment for Ebola virus infection, a virus-like particle (VLP) vaccine is effective in mice, guinea pigs, and non-human primates when given pre-infection. In this work, we report that VLPs protect Ebola virus-infected mice when given 24 hours post-infection. Analysis of cytokine expression in serum revealed a decrease in pro-inflammatory cytokine and chemokine levels in mice given VLPs post-exposure compared to infected, untreated mice. Using knockout mice, we show that VLP-mediated post-exposure protection requires perforin, B cells, macrophages, conventional dendritic cells (cDCs), and either CD4+ or CD8+ T cells. Protection was Ebola virus-specific, as marburgvirus VLPs did not protect Ebola virus-infected mice. Increased antibody production in VLP-treated mice correlated with protection, and macrophages were required for this increased production. However, NK cells, IFN-gamma, and TNF-alpha were not required for post-exposure-mediated protection. These data suggest that a non-replicating Ebola virus vaccine can provide post-exposure protection and that the mechanisms of immune protection in this setting require both increased antibody production and generation of cytotoxic T cells.

Controlled analysis of nanoparticle charge on mucosal and systemic antibody responses following pulmonary immunization

Pulmonary immunization enhances local humoral and cell-mediated mucosal protection, which are critical for vaccination against lung-specific pathogens such as influenza or tuberculosis. A variety of nanoparticle (NP) formulations have been tested preclinically for pulmonary vaccine development, yet the role of NP surface charge on downstream immune responses remains poorly understood. We used the Particle Replication in Non-Wetting Templates (PRINT) process to synthesize hydrogel NPs that varied only in surface charge and otherwise maintained constant size, shape, and antigen loading. Pulmonary immunization with ovalbumin (OVA)-conjugated cationic NPs led to enhanced systemic and lung antibody titers compared with anionic NPs. Increased antibody production correlated with robust germinal center B-cell expansion and increased activated CD4(+) T-cell populations in lung draining lymph nodes. Ex vivo treatment of dendritic cells (DCs) with OVA-conjugated cationic NPs induced robust antigen-specific T-cell proliferation with ∼ 100-fold more potency than soluble OVA alone. Enhanced T-cell expansion correlated with increased expression of surface MHCII, T-cell coactivating receptors, and key cytokines/chemokine expression by DCs treated with cationic NPs, which were not observed with anionic NPs or soluble OVA. Together, these studies highlight the importance of NP surface charge when designing pulmonary vaccines, and our findings support the notion that cationic NP platforms engender potent humoral and mucosal immune responses.

Oestrogen up‐regulates interleukin‐21 production by CD4+T lymphocytes in patients with systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is an autoimmune disease in which abnormal immune responses are mediated by tissue-binding autoantibodies and immune complex deposition. Because most SLE patients are women of child-bearing age, oestrogen has been suggested to play an important role in SLE pathogenesis. One proposed role is to induce B-cell activation, culminating in increased autoantibody production. Interleukin-21 (IL-21) has been shown to be crucial in the differentiation of activated B cells into plasma cells. We therefore hypothesized that oestrogen up-regulates IL-21 production and induces subsequent B-cell activation in SLE patients. Peripheral blood was obtained from 22 SLE patients and 16 healthy controls. Expression levels of IL-21 and its receptor in serum, peripheral blood mononuclear cells, and CD4(+) T cells were higher in SLE patients than in healthy controls. Exposure of CD4(+) T cells from SLE patients to 17β-oestradiol led to a dose- and time-dependent increase in IL-21 expression, which was abolished in the presence of mitogen-activated protein kinase (MAPK) (MAPK kinase, p38, Jun N-terminal kinase) inhibitors. B cells from healthy controls showed increased antibody production when they were co-cultured with oestrogen-treated CD4(+) T cells from SLE patients. Treatment with IL-21 antibody abrogated the increased antibody production of the co-culture systems. This study revealed the association between oestrogen and IL-21 in SLE patients. Oestrogen up-regulates IL-21 expression of CD4(+) T cells via MAPK-dependent pathways in SLE patients, which in turn induces increased antibody production by B cells.

THU0532 Estrogen Upregulates Interleukin-21 Production of Clusters of Differentiation 4 Positive T Lymphocytes in Patients with Systemic Lupus Erythematosus

BackgroundSystemic lupus erythematosus (SLE) is an autoimmune disease in which various organs and tissues are damaged through abnormal immune responses mediated by tissue-binding autoantibodies and immune complex deposition. As the...

Mekabu fucoidan: Structural complexity and defensive effects against avian influenza A viruses

Fucoidan from the sporophyll (Mekabu) of brown seaweed Undaria pinnatifida (wakame) is interesting due to its various biological activities. Mekabu fucoidan (Mw ∼9kDa) of this study (MF) was previously isolated and characterized by chemical and separation methods including GPC and methylation analysis (Lee, Hayashi, Hashimoto, Nakano, & Hayashi, 2004). It was found that this fucoidan composed of partially sulphated (DS ∼0.72) fucose and galactose at approximately equal amounts. Methylation analyses revealed complex structure of MF. However, it has been still unclear about the linkages between units and substitution patterns. To solve these structural tasks, spectroscopic methods (FTIR, FT Raman and NMR) were used in the analysis of native MF and its deesterified derivatives. According to obtained results, this polysaccharide was defined as O-acetylated sulphated fucogalactan. The defensive effects of MF were evaluated on mice infected with avian influenza A viruses (H5N3 and H7N2 subtypes); its efficacy was determined in reducing viral replication and increasing antibody production. Oral administration of MF resulted in suppressing virus yields. In addition, the production of neutralizing antibodies and mucosal IgA in the animals inoculated with the avian influenza A viruses was significantly increased. These results suggested that MF could be used for the prevention of viral infection.

Applications of low-intensity pulsed ultrasound to increase monoclonal antibody production in CHO cells using shake flasks or wavebags

Many technologies, such as cell line screening and host cell engineering, culture media optimization and bioprocess optimization, have been proposed to increase monoclonal antibody (mAb) production in Chinese Hamster Ovary (CHO) cells. Unlike the existing biochemical approaches, we investigated stimulation using low-intensity pulsed ultrasound (LIPUS) as a purely physical approach, offering enhanced scalability, contamination control and cost-efficiency, while demonstrating significantly increased cell growth and antibody production. It was found that daily ultrasound treatments at 40 mW/cm2 for 5min during cell culture increased the production of human anti-IL-8 antibody by more than 30% using 10 or 30mL shake flasks. Further increasing the ultrasound dosage (either intensities or the treatment duration) did not appreciably increase cell growth or antibody production, however feeding the culture with additional highly-concentrated nutrients, glucose and amino acids (glutamine in this case), did further increase cell growth and antibody titer to 35%. Similar ultrasound treatments (40 mW/cm2, 5min per day) when scaled up to larger volume wavebags, resulted in a 25% increase in antibody production. Increased antibody production can be attributed to both elevated cell count and the ultrasound stimulation. Theoretical study of underlying mechanisms was performed through the simulations of molecular dynamics using the AMBER software package, with results showing that LIPUS increases cell permeability. The significance of this study is that LIPUS, as a physical-based stimulation approach, can be externally applied to the cell culture without worrying about contamination. By combining with the existing technologies in antibody production, LIPUS can achieve additional mAb yields. Because it can be easily integrated with existing cell culture apparatuses, the technology is expected to be more acceptable by the end users.

Vaccination with DHA-derived specialized proresolving mediators increases the antibody-mediated immune response against influenza virus (VAC2P.925)

Abstract Influenza viruses remain a critical global health concern. More efficacious vaccines are needed to protect against influenza virus, yet few adjuvants are approved for routine use. Specialized proresolving mediators (SPMs) are powerful endogenous bioactive lipid regulators of inflammation, with great clinical translational properties. We previously identified endogenous levels of the SPM resolvin D1, protectin D1 and 17-hydroxydocosahexaenoic acid (17-HDHA) in the mouse spleen. Here, we investigated the ability of 17-HDHA to enhance the adaptive immune response using a pre-clinical influenza vaccination mouse model. Our findings revealed that mice immunized with influenza H1N1-derived HA protein plus 17-HDHA increased antigen specific antibody titers. 17-HDHA increased the number of plasma cells and HA-specific antibody-secreting cells in the bone marrow, but it did not affect the frequency of GC B cells or Tfh cells in the draining lymph node. Importantly, the 17-HDHA-mediated increased antibody production was more protective against live pH1N1 influenza virus infection in mice. This is the first report on the immuno-stimulatory effects of omega-3-derived SPMs on the humoral immune response. These findings illustrate a previously unknown biological relation between proresolution signals and the adaptive immune system. In addition, 17-HDHA as well as other immuno-stimulatory SPM have the potential to be used as novel endogenous adjuvants.

α-Galactosylceramide stimulates splenic lymphocyte proliferation in vitro and increases antibody production in vivo in the late neonatal-age mouse. (VAC12P.1028)

Abstract The neonatal stage is characterized by weak responses to various infections and vaccines; thus the development of efficient formulas to improve vaccine effectiveness is of high priority. The glycolipid alpha galactosylceramide (αGalCer) is known as a potent immune modulator mainly due to NKT cell activation. Using a mouse tetanus toxoid (TT) immunization model, we observed that neonatal mice given αGalCer at the time of primary immunization on day (d) 17 had a significant higher TT-specific IgM response (ELISA), while given αGalCer at d7 during immunization only resulted in a marginal boosting. Consistently, immunostaining of the spleen sections from αGalCer-treated d17 immunized neonates showed higher number of Ki67+ cells in the splenic germinal center area, suggesting stronger response after immunization. In vitro kinetic studies revealed that spleen cells from newborn to d7 neonates did not respond to αGalCer stimulation, whereas cell proliferation was markedly increased by αGalCer after d7, and became dramatic around neonatal d17-18, which was accompanied by increased B and T lymphocyte populations in the spleen. In addition, in d17 spleen cells, αGalCer significantly stimulated the proliferation of CD23+ B cells, a subset of B cells enriched in germinal centers. These data suggest that αGalCer is an effective stimulator of the immune system in late neonatal stage and thus may be useful as a potential adjuvant to achieve an efficient immunization.

EPA and DHA increase the frequency of murine B cell subsets and restore antibody production in obesity upon stimulation with a T‐independent antigen (382.5)

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), the bioactive components of fish oil, exert immunomodulatory properties. The impact of EPA and DHA supplementation on murine B cell function is poorly studied. We first determined the effects of fish oil on B cell phenotypes and antibody production in obese mice upon stimulation with a T‐independent antigen. Fish oil intervention increased the frequency of transitional 1 and marginal zone B cells in obese mice. Fish oil restored the decrement in circulating IgM in obesity and increased antibody production in lean mice. We then measured the effects of EPA and DHA ethyl esters on B cell activation upon supplementation to an obesogenic diet. Ex vivo stimulation of B cells with lipopolysaccharide increased B cell activation as measured by TNF‐α and IL‐6 secretion. DHA, but not EPA, increased B cell IL‐10 production. In vivo, EPA and DHA differentially increased the frequency of transitional 1, transitional 2 and pre‐marginal zone cells. Taken together, the data establish that fish oil can boost B‐cell mediated immunity in obesity and that EPA and DHA ethyl esters exert differential effects on B cells.

Interrelationships between fish nutrition and health

An understanding of how to nurture and/or modulate the different components of the immune system is crucial for the prevention and control of diseases in animal husbandry. It is well established that proper nutrition is essential for maintenance of normal growth and health of all animals including aquatic species. As such, nutritious diets and appropriate feeding regimes play critical roles in intensive aquaculture. In recent years, heightened attention has been given to the development of nutritional strategies that positively influence immunity and disease resistance of cultured organisms to reduce disease-related economic losses. The availability of specific nutrients to immune cells plays a key role on how well those cells perform against a foreign invader. In this sense, research with several fish species has established that not only immunocompetence can be compromised by deficiencies of various nutrients, but that dietary supplementation of some nutrients in excess of minimum requirement levels for optimal growth has been shown to significantly enhance immune responses and disease resistance. For instance, findings from recent studies indicate an important role of key amino acids and their derivatives, like arginine and glutamine, or vitamins, such as vitamin C and E, in modulating immune responses such as increasing phagocytosis and pathogen killing capacity, as well as increasing antibody production and immunological memory. In addition, administration of non-nutritive compounds in the diet has become recognized as a viable means of enhancing immunocompetence of various aquatic species, mainly by the provision of cellular fractions such as β-glucans, complex oligosaccharides (mannanoligosaccharides, fructooligosaccharides or sulfated polysaccharides) or yeast and algae extracts. These compounds may act like Pathogen Associated Molecular Pattern (PAMP) molecules, interacting with the innate immune system through their Pattern Recognition Receptors (PRRs) and increasing their capability for detecting and recognizing potential pathogens and readily triggering immune responses. This article will review the broad range of dietary constituents which have been shown to affect immunocompetence and health of aquatic species and how they may influence components of the immune system. Further advancements in nutritional modulation of the immune response hold promise as an effective and relatively inexpensive alternative in combating diseases in aquaculture.

Antibodies are involved in the protective immunity induced in mice by Schistosoma mansoni schistosomula tegument (Smteg) immunization

The Schistosoma mansoni schistosomula tegument (Smteg) plays an important role in triggering the host immune response and mice immunization with Smteg formulated with Freunds adjuvant or alum + CpG induce partial protection against S. mansoni infection associated with an increased antibody production. In this study, we investigated the role of these antibodies in parasite killing both in vitro and in vivo. We demonstrated that these antibodies were able to bind to the surface of S. mansoni recently transformed schistosomula and that these antibodies significantly increase the percentage of schistosomula killed in vitro by complement activation. Passive transference of immune sera decreased the parasite burden and the number of eggs trapped in the organs of mice that received sera containing anti-Smteg antibodies. These results demonstrate that antibodies specific to surface tegumental antigens are involved in parasite elimination in mice immunized with Smteg.

Pemphigus vulgaris in pregnancy associated with herpes virus type 1 infection.

Dear Editor: Pemphigus vulgaris (PV) rarely occurs during pregnancy. We report a case of PV associated with herpes simplex type 1 virus (HSV-1) which occurred in the third trimester of pregnancy. A 32-year-old second gravida at 37 weeks' gestation was admitted for multiple bullous skin lesions that persisted for over a month. These vesicular and erosive lesions initiated from the periumbilical region and spread to the oral mucosa and the skin of the back (Fig. 1A, B). The diagnosis of PV was confirmed by biopsy (Fig. 1C), and direct immufluorescence detected anti-immunoglobulin G and C3 antibodies. Anti-desmoglein 1 and anti-desmoglein 3 antibodies were elevated at 82.1 U/ml (normal, <14 U/ml) and 184.9 U/ml (normal, <7 U/ml) in peripheral blood. Tzanck smear and viral polymerase chain reaction (Seeplex STD B41 Detedtion; Seegene, Seoul, Korea) were done on the base of a vesicular lesion on the trunk. Tzanck smear was negative, but, viral polymerase chain reaction (PCR) was positive for HSV-1 (Fig. 2). Fig. 1 (A) Multiple erosions on oral mucosa and (B) scattered crusted vesicles and erosions on the back. (C) Suprabasal blisters with eosinophils and acantholytic cells in the epidermis (H&E, ×400). Fig. 2 Polymerase chain reaction results show a positive band on 189 bp. M: 100 bp DNA ladder, Lane 1: positive result of the patient's tissue for herpes simplex type 1 virus (189 bp), Lane 2: normal control, Lane 3: positive control, Lane 4: sensitivity test, ... Prednisolone at a dose of 20 mg/d was initiated. Following PCR results, antiviral agent (valacyclovir, total 4,500 mg) was also given for 5 days. Lesions started to improve after starting the medication. Two weeks after the diagnosis the patient was transferred to the labour unit for induced labour. The baby was in a good state. After giving birth, the dose of prednisolone was increased to 25 mg/d, and mycophenolate mofetil 1.0 g/d was added. After improvement in both vesicles and general health status, the patient was discharged. During follow-up, a subtle amount of skin lesion occurred which was controlled with topical steroid, oral mycofenolate mofetil 1.0 g/d, and intermittent oral steroid at a dose of 4 mg/d. Though it is known that pemphigus develops as a result of the interaction between endogenous (genetic) factors and exogenous factors1, the primary stimulus for the autoimmune response of PV is still unknown. Pregnancy may aggravate certain autoimmune diseases, such as systemic lupus erythematosus, pemphigoid gestationis, and PV2. The patient's vesicular eruptions developed during the period of conception and were not controlled well at that time, so we supposed her condition was related to pregnancy. Bullous diseases that are reported to be associated with pregnancy include pemphigoid gestationis pemphigus foliaceus, and PV2,3. The most important diagnostic tool is histologic examination, and the two diseases mentioned above show subepidermal, subcorneal blisters. However PV, as seen in our patient, shows a suprabasal epidermal split with acantholysis and few eosinophils in the dermis, and perilesional skin shows intercelluar deposits of IgG and C3 in the epidermis on direct immunofluorescence4. A diagnosis of PV in this patient was made according to the histological features and clinical involvement of oral mucosa. The possible role of viruses, especially HSV-1, has been proposed in the pathogenesis of PV. Positive viral PCR results suggest that viral factors, especially HSV and Epstein-Barr virus, are capable of inducing and/or exacerbating pemphigus in a genetically susceptible host1. Infectious agents can stimulate the immune response in genetically susceptible individuals or in those with immune deviating conditions such as pregnancy, leading to an increase in the production of cytokines. High levels of interferon-gamma induce the expression of human leucocyte antigen type 2 in the membranes of keratinocytes, making the structural site of PV antigen (epitope spreading) immunologically active. Chronic, recurrent viral infections can also stimulate excessive production of interleukin (IL)-4 and IL-10, which result in a shift from TH1 to TH2 response, increasing antibody production. In addition, they can also directly infect B and T lymphocytes, contributing to the production of autoreactive B lymphocytes and autoimmune antibodies5. When infected, keratinocytes can pass through structural changes which favor the exposure of antigens1. In summary, we report a case of a woman who showed PV associated with HSV-1 in her second pregnancy, which was cured with steroids and antiviral agents.

Autoimmune Disease pH and Temperature.

Perhaps parts of the immune system are pH and temperature sensitive. If true, there could be a variance in pH and/or temperature under certain conditions that could impact the immune system and vice versa. Possibly the illusive etiology of autoimmune disease is in part due to pH and/or temperature changes. A majority would probably agree the human immune system is designed to destroy foreign invasion such as an infection and/or for removal of nonviable tissue such as that damaged by trauma. Obviously, both of the previous situations would be more likely to involve at least a local environment trending more acidic. Also most would agree autoimmune disease is when there is an error in the immune system and the system begins to attack and destroy viable tissue. Thus could the etiology of autoimmune disease be a repair system gone awry possibly because of a pH and/ or temperature change? If cancer is the repair system gone awry in regard to overproduction of replacement cells, then perhaps autoimmune disease is the repair system gone awry in regard to the damaged cell removal causing disease possibly because of a pH and/or temperature change. When there is cell damage inflammation develops as certain cells infiltrate the area and certain mediators accumulate that are also part of the immune system. Autoantibodies have been noted to be present following trauma with some suspecting this is a mechanism to remove cellular debris. However, autoantibody presence suggests a possible autoimmune disease in the future but is not diagnostic, since many normal individuals have mild levels. For example, with diabetes mellitus type I, there can be an autoantibody present for years (or even a life time) without any overt disease ever developing. In short, autoimmunity may follow tissue damage with low levels of antibodies but not necessarily actual autoimmune disease with destruction of viable tissue. So this is similar to other situations where something could occur, but does not have to occur, or rather something is more likely to occur, but is still not a certainty. An unknown second step probably has to occur, could that second step involve a pH and/or temperature change is the question. If an individual develops an autoimmune disease, they are more likely to develop a second autoimmune disease than are other patients. Autoimmune diseases seem to run in families as well. There appears to be a genetic susceptibility. Some say autoimmune diseases are multigenic, heterogeneous in genetic basis, with identical twins mixed on penetrance but usually higher than siblings and much higher than unrelated populations. If there is a pH aspect to the immune system, perhaps this involves the cellular pumps, channels, transporters and isoenzyme levels. This could account for a familial predisposition yet be multigenic, since there are so many different cellular pumps, channels, transporters and isoenzymes. There are multiples of each of these, so a particular combination could make an individual susceptible and this would be genetic. The cellular pumps, channels, transporters and isoenzymes could also respond to environmental stress changing to some degree accounting for differences in identical twins. The medications procainamide and hydralazine have been known for decades to induce autoimmune disease, specifically lupus. Procainamide impacts sodium channels while hydralazine impacts the calcium activated potassium channels and so both could impact pH levels. Interestingly, some compliment activation [1] and IgG binding [2] are more reactive in an acidic environment. In addition, some interleukins seem to prefer an acidic environment [3]. But there is also a possibility extreme levels of alkalinity can set the immune system into action as well, at the very least through cell destruction. Strong ions such as iodine in the thyroid could conceivably impact the cellular pumps, channels, transporters and isoenzymes. Indeed that is possibly what we see. As the level of intrathyroidal iodine increases, so does the incident of Hashimoto’s thyroiditis [4], one of the most common autoimmune diseases. However, not everyone develops Hashimoto’s thyroiditis with higher iodine, just some people. Could those with a certain mix of cellular pumps, channels, transporters and isoenzymes be the ones that are more likely to develop the Hashimoto’s thyroiditis? Also some cytokines such as interferon can induce Hashimoto’s thyroiditis. Could interferon possibly be impacting the cellular pumps, channels, transporters and isoenzymes? Yes, it is known to impact the potassium channels for example [5]. Finally, if pH changes enough there could be a charge generated which could impact movement possibly even for T and B cells. Another interesting aspect about autoimmune diseases is they are in general significantly more common usually in females than males. Women have increased antibody production compared to Manuscript accepted for publication April 10, 2014

Proteins improving recombinant antibody production in mammalian cells

Mammalian cells have been successfully used for the industrial manufacture of antibodies due to their ability to synthesize antibodies correctly. Nascent polypeptides must be subjected to protein folding and assembly in the ER and the Golgi to be secreted as mature proteins. If these reactions do not proceed appropriately, unfolded or misfolded proteins are degraded by the ER-associated degradation (ERAD) pathway. The accumulation of unfolded proteins or intracellular antibody crystals accompanied by this failure triggers the unfolded protein response (UPR), which can considerably attenuate the levels of translation, folding, assembly, and secretion, resulting in reduction of antibody productivity. Accumulating studies by omics-based analysis of recombinant mammalian cells suggest that not only protein secretion processes including protein folding and assembly but also translation are likely to be the rate-limiting factors for increasing antibody production. Here, this review describes the mechanism of antibody folding and assembly and recent advantages which could improve recombinant antibody production in mammalian cells by utilizing proteins such as ER chaperones or UPR-related proteins.

Supplementation of Elderly Japanese Men and Women with Fucoidan from Seaweed Increases Immune Responses to Seasonal Influenza Vaccination

The elderly are known to have an inadequate immune response to influenza vaccine. Mekabu fucoidan (MF), a sulfated polysaccharide extracted from seaweed, was previously shown to have an immunomodulatory effect. We therefore investigated antibody production after influenza vaccination in elderly Japanese men and women with and without oral MF intake. A randomized, placebo-controlled, double-blind study was conducted with 70 volunteers >60 y of age. They were randomly assigned to 1 of 2 groups, consuming either MF (300 mg/d) or placebo for 4 wk, and then given a trivalent seasonal influenza vaccine. Serum was sampled at 5 and 20 wk after vaccination to measure the hemagglutination inhibition titer and natural killer cell activity. The MF group had higher antibody titers against all 3 strains contained in the seasonal influenza virus vaccine than the placebo group. Titers against the B/Brisbane/60/2008 (B) strain increased substantially more in the MF group than in the placebo group over the product consumption period. The immune response against B antigen met the European Union Licensure criteria regarding the geometric mean titer ratio in the MF group (2.4), but not in the placebo group (1.7). In the MF group, natural killer cell activity tended to increase from baseline 9 wk after MF intake (P = 0.08). However, in the placebo group no substantial increase was noted at 9 wk, and the activity decreased substantially from 9 to 24 wk. In the immunocompromised elderly, MF intake increased antibody production after vaccination, possibly preventing influenza epidemics.

Enhancement of monoclonal antibody production in CHO cells by exposure to He-Ne laser radiation.

This study tested the effectiveness of laser biostimulation in small-scale cultures in vitro. We investigated the response of recombinant CHO cells, which are used for the production of monoclonal antibody, to low level laser radiation. The cells were irradiated using a 632.8nm He-Ne laser in a continuous wave mode at different energy doses. We incubated the irradiated cells in small batch cultures and assessed their proliferation and productivity at various time intervals. Compared to untreated cells, the irradiated cells showed a significant increase in antibody production. Moreover, the results showed that laser irradiation did not affect viability and slightly enhanced proliferation rate.

AB0203 The role of estrogens on the production of IL-21 in patients with systemic lupus erythematosus

BackgroundSystemic lupus erythematosus (SLE) is an autoimmune disease in which various organs and tissues are damaged through abnormal immune responses mediated by tissue-binding autoantibodies. Though, the exact pathogenic mechanism is...