Karl A. Brokstad

Influenza Virus: Immunity and Vaccination Strategies. Comparison of the Immune Response to Inactivated and Live, Attenuated Influenza Vaccines

Influenza virus is a globally important respiratory pathogen which causes a high degree of morbidity and mortality annually. The virus is continuously undergoing antigenic change and thus bypasses the hosts acquired immunity to influenza. Despite the improvement in antiviral therapy during the last decade, vaccination is still the most effective method of prophylaxis. Vaccination induces a good degree of protection (60–90% efficacy) and is well tolerated by the recipient. For those at risk of complications from influenza, annual vaccination is recommended due to the antigenic changes in circulating strains. However, there is still room for improvement in vaccine efficacy, long‐lasting effect, ease of administration and compliance rates. The mucosal tissues of the respiratory tract are the main portal entry of influenza, and the mucosal immune system provides the first line of defence against infection. Secretory immunoglobulin A (SIgA) and IgM are the major neutralizing antibodies directed against mucosal pathogens. These antibodies work to prevent pathogen entry and can function intracellularly to inhibit replication of virus. This review describes influenza virus infection, epidemiology, clinical presentation and immune system response, particularly as it pertains to mucosal immunity and vaccine use. Specifically, this review provides an update of the current status on influenza vaccination and concentrates on the two main types of influenza vaccines currently in use, namely the cold‐adapted vaccine (CAV) given intranasally/orally, and the inactivated vaccine (IV) delivered subcutanously or intramuscularly. The commercially available trivalent IV (TIV) elicits good serum antibody responses but induces poorly mucosal IgA antibody and cell‐mediated immunity. In contrast, the CAV may elicit a long‐lasting, broader immune (humoral and cellular) response, which more closely resembles natural immunity. The immune response induced by these two vaccines will be compared in this review.

Neuropsychiatric disturbances in SLE are associated with antibodies against NMDA receptors

To determine whether neuropsychiatric manifestations in patients with systemic lupus erythematosus (SLE) are influenced by antibodies against the human N‐methyl‐d‐aspartate (NMDA) receptor types NR2a or NR2b. A decapeptide was synthesized containing a sequence motif present in the extracellular ligand‐binding domain of NMDA receptors NR2a and NR2b, bound by the monoclonal murine anti‐DNA antibody R4A. In an ELISA with the murine monoclonal R4v as positive control, plasma samples of 57 patients with SLE were examined for the anti‐peptide (anti‐NR2) antibody after the patients had been subjected to comprehensive psychological and cognitive testing. Poor performance on the Visual Paired Associates test (immediate), the Grooved Pegboard test, as well as high scores on the Beck Depression Inventory, and scales D‐2 (depression), Pd‐4 (psychopathic deviate), Sc‐8 (schizophrenia), and Ma‐9 (hypomania) of the MMPI‐2 were significantly associated with elevated levels of anti‐NR2 antibodies. The findings in several domains indicate an association between anti‐NR2 antibodies and depressed mood in addition to decreased short‐time memory and learning. Antibodies to NMDA receptors thus may represent one of several mechanisms for cerebral dysfunction in patients with SLE.

The human immunodeficiency virus type 1 Rev protein shuttles between the cytoplasm and nuclear compartments.

A retroviral regulatory protein, Rev (regulator of virion protein expression), is made in cells infected by human immunodeficiency virus (HIV). Rev is essential for the completion of the retroviral life cycle and interacts with the host cell at some posttranscriptional step in order to express the incompletely spliced HIV mRNAs from which HIV structural proteins are translated. Neither the host cell components nor the mechanisms responsible for this important regulation have been defined. We now report that Rev is a nucleocytoplasmic shuttle protein which is continuously transported between the cytoplasm, the nucleoli, and nucleoplasmic speckles enriched in RNA splicing and processing factors. The results show that Rev has the potential to interfere specifically with the splicing of the HIV pre-mRNA in the nucleoplasm and, next, guide such mRNAs to the cytoplasm for translation.

Subcellular redistribution and surface exposure of the Ro52, Ro60 and La48 autoantigens during apoptosis in human ductal epithelial cells: a possible mechanism in the pathogenesis of Sjögren's syndrome.

The Ro52, Ro60 and La48 autoantigens are associated with Sjögrens syndrome (SS) and systemic lupus erythematosus (SLE). The mechanisms behind tolerance breakdown of these self‐peptides remain unclear; however, apoptosis has been proposed to cause their presentation to the immune system. We have examined the localization of transiently expressed enhanced green fluorescent protein (EGFP)‐tagged Ro52, Ro60 and La48 autoantigens in a human salivary gland (HSG) cell line by laser confocal microscopy under normal growth conditions and during apoptosis. Surface exposure of Ro52, Ro60 and La48 was demonstrated on nonfixed apoptotic cells with monoclonal antibodies (MoAbs) or with primary SS patient antisera. Laser scanning cytometry determined the apoptotic frequency. EGFP alone was studied as control. We found that Ro52 mainly is cytoplasmic, Ro60 both nuclear and cytoplasmic, while La48 only resides in the nucleus under normal conditions. During early apoptosis, La48 is dramatically redistributed to the cytoplasm, while the localization of Ro52 and Ro60 is maintained. All three autoantigens filled apoptotic blebs and covered TUNEL (terminal‐deoxynucleotidyl‐transferase‐mediated dUTP–digoxigenin nick end labelling)‐positive apoptotic bodies. Identical results were obtained in COS‐7 cells. We have developed a transfection system to study the intracellular localization of the three autoantigens Ro52, Ro60 and La48, without antibody detection. During apoptosis, there is an intracellular redistribution of endogenous and EGFP‐tagged Ro52, Ro60 and La48, leading to surface exposure. These findings may indicate a role for apoptosis in the induction and facilitation of humoral responses to Ro52, Ro60 and La48 in the autoimmune exocrinopathy of SS.

Autoantibodies Present Before Symptom Onset in Primary Sjögren Syndrome

Methods | All patients with primary Sjögren syndrome at Malmö UniversityHospital(Malmö,Sweden)havebeenincludedinaregistry since 1984.3 To obtain presymptomatic serum samples, the registry was linked with 3 biobanks containing specimens from 625 000 individuals submitted for microbiological analyses or population-based studies of healthy individuals.4 Date of symptom onset was determined retrospectively from the patient during the first office visit at which Sjögren syndrome was diagnosed. All cases provided written informed consent at inclusion in the registry. The study was approved by the ethics committee at Lund University. All patients with available samples who met consensus criteria for Sjögren syndrome1 were included. When multiple samples were available for a single patient, the earliest positive sample was used. Controls were randomly selected from the biobanks and matched by sex, age, and date of earliest sampling (within 60 days before or after) to each case. None of the controls were diagnosed with Sjögren syndrome. Those serum samples that were obtained from the microbiology biobank had been submitted due to symptoms unrelated to Sjögren syndrome (eg, pregnancy screening, suspected influenza). Samples were collected from 1976 through 2001 and Sjögren syndrome was diagnosed through 2011. Autoantibodies against Ro60/SSA, Ro52/SSA, La/SSB, Sm, RNP, Scl-70, Jo-1, ribosome P, and chromatin were detected using a multiplex immunobead assay (QUANTA Plex SLE Profile 8, INOVA Diagnostics) and analyzed on a Luminex 100 instrument (Luminex Corp). Antinuclear antibodies (ANAs) were analyzed by immunofluorescence with HEp-2 cells as the antigens. Immunoglobulin M-class rheumatoid factor (RF) was analyzed with an in-house enzyme-linked immunosorbent assay. Descriptive statistics and a 2-sided Friedman test were used for statistical analysis (Statistics version 20.0; IBM SPSS). P < .05 was considered statistically significant.

Sjögren's Syndrome—A Plethora of Clinical and Immunological Phenotypes with a Complex Genetic Background

Abstract:  Primary Sjögrens syndrome is a complex autoimmune disorder, considered to represent an ideal disease with which to study the mechanisms underlying autoimmunity because its manifestations are both organ specific and systemic in nature. The characteristic histologic finding in target organs is a progressive focal infiltration of mononuclear lymphoid cells, replacing glandular epithelium (lymphoepithelial lesion). This involvement has been re‐emphasized in the 2002 revised EU criteria for Sjögrens syndrome. Moreover, ectopic secondary lymphoid follicles in Sjögrens syndrome contain all elements of relevance for driving an autoimmune response. A number of cytokines and chemokines are involved and particularly B cell activating factor seems to direct the lifespan of infiltrating B cells by enhancing their proliferation and maturation. The recent discovery of clinical benefit after B cell depletion also highlights the pivotal role of B cells in Sjögrens syndrome. A major challenge in Sjögrens syndrome will be to stratify the disease process including genetic and environmental triggers. Identification of novel genetic and molecular markers may lead to the development of better diagnostic and prognostic tools in Sjögrens syndrome including its systemic complications. This minor review will cover the current knowledge on classification, pathogenesis, multiplex findings, potential candidate genes, gene profiling results, and novel therapy approaches. New hypotheses behind the complexity of Sjögrens syndrome are expected to follow.

Salivary glands of primary Sjögren's syndrome patients express factors vital for plasma cell survival

IntroductionThe presence of circulating Ro/SSA and La/SSB autoantibodies has become an important marker in the classification criteria for primary Sjögrens syndrome (pSS). Plasma cells producing these autoantibodies are mainly high affinity plasma cells originating from germinal centre reactions. When exposed to the right microenvironment these autoimmune plasma cells become long-lived and resistant to immunosuppressive treatment. Since autoimmune plasma cells have been detected in the salivary glands of SS patients, we wanted to investigate if the glandular microenvironment is suitable for plasma cell survival and if glandular residing plasma cells are the long-lived plasma cell subset.MethodsSingle, double and triple immunohistochemistry as well as immunofluorescence staining was performed on minor salivary gland tissue retrieved from pSS, chronically inflamed and normal subjects.ResultsWe detected significant numbers of CD138+, non-proliferating, Bcl-2 expressing plasma cells in the salivary glands of pSS patients with high focus score (FS). Furthermore, we demonstrated that CXCL12 and interleukin (IL)-6 survival factors were highly expressed in pSS salivary gland epithelium and by focal mononuclear infiltrating cells. Notably, adipocytes when present in the salivary gland tissue were an important source of CXCL12. We clearly demonstrate that plasma cells are localised in close proximity to CXCL12 and IL-6 expressing cells and thus that the environment of salivary glands with high FS provide factors vital for plasma cell survival.ConclusionsPlasma cells residing in the salivary glands of pSS patients with high FS showed phenotypic characteristics of the long-lived plasma cell subtype. Furthermore, the pSS salivary gland microenvironment provided niches rich in factors vital for plasma cell survival.

Influenza Vaccination Strategies: Comparing Inactivated and Live Attenuated Influenza Vaccines

Influenza is a major respiratory pathogen causing annual outbreaks and occasional pandemics. Influenza vaccination is the major method of prophylaxis. Currently annual influenza vaccination is recommended for groups at high risk of complications from influenza infection such as pregnant women, young children, people with underlying disease and the elderly, along with occupational groups such a healthcare workers and farm workers. There are two main types of vaccines available: the parenteral inactivated influenza vaccine and the intranasal live attenuated influenza vaccine. The inactivated vaccines are licensed from 6 months of age and have been used for more than 50 years with a good safety profile. Inactivated vaccines are standardized according to the presence of the viral major surface glycoprotein hemagglutinin and protection is mediated by the induction of vaccine strain specific antibody responses. In contrast, the live attenuated vaccines are licensed in Europe for children from 2–17 years of age and provide a multifaceted immune response with local and systemic antibody and T cell responses but with no clear correlate of protection. Here we discuss the immunological immune responses elicited by the two vaccines and discuss future work to better define correlates of protection.

Prediction of Sjögren's Syndrome Years Before Diagnosis and Identification of Patients With Early Onset and Severe Disease Course by Autoantibody Profiling.

Autoantibodies are highly characteristic of primary Sjögrens syndrome (SS) and represent important tools for studying its pathogenesis. Nonetheless, thus far, no systematic investigations have assessed the presence of autoantibodies before diagnosis. This study was undertaken to analyze how early and in what order autoantibodies appear, how predictive they are of primary SS, and whether they identify disease subsets.

Parenteral Vaccination against Influenza Does Not Induce a Local Antigen-Specific Immune Response in the Nasal Mucosa

The immune response in the nasal mucosa to influenza vaccination in 23 patients scheduled for tonsillectomy was studied. A statistically significant increase in influenza virus-specific serum and oral fluid antibodies was observed 7 days after vaccination. The numbers of influenza virus-specific antibody-secreting cells (ASCs) in peripheral blood also increased significantly 1 week after vaccination. The numbers of ASCs in tonsils and nasal mucosa were compared with data from a recent study of nonvaccinated volunteers. The numbers of influenza virus-specific ASCs in tonsils were significantly higher in the vaccinated group, but, surprisingly, there was no significant difference between the groups in the numbers of ASCs in nasal mucosa. This suggests that the influenza virus-specific antibodies detected in oral fluid are not produced locally in the nasal mucosa and may originate from a systemic source, indicating that the vaccination may favor a systemic immune response.