Meningococcal omp85 in detergent-extracted outer membrane vesicle vaccines induces high levels of non-functional antibodies in mice.

The vaccine potential of meningococcal Omp85 was studied by comparing the immune responses of genetically modified deoxycholate-extracted outer membrane vesicles, expressing five-fold higher levels of Omp85, with wild-type vesicles. Groups (n = 6-12) of inbred and outbred mouse strains (Balb/c, C57BL/6, OFI and NMRI) were immunized with the two vaccines, and the induced antibody levels and bactericidal and opsonic activities measured. Except for Balb/c mice, which were low responders, the genetically modified vaccine raised high Omp85 antibody levels in all mouse strains. In comparison, the wild-type vaccine gave lower antibody levels, but NMRI mice responded to this vaccine with the same high levels as the modified vaccine in the other strains. Although the vaccines induced strain-dependent Omp85 antibody responses, the mouse strains showed high and similar serum bactericidal titres. Titres were negligible with heterologous or PorA-negative meningococcal target strains, demonstrating the presence of the dominant bactericidal PorA antibodies. The two vaccines induced the same opsonic titres. Thus, the genetically modified vaccine with high Omp85 antibody levels and the wild-type vaccine induced the same levels of functional activities related to protection against meningococcal disease, suggesting that meningococcal Omp85 is a less attractive vaccine antigen.

Salivary IgA from the sublingual compartment as a novel noninvasive proxy for intestinal immune induction.

Whole-saliva IgA appears like an attractive noninvasive readout for intestinal immune induction after enteric infection or vaccination, but has failed to show consistent correlation with established invasive markers and IgA in feces or intestinal lavage. For reference, we measured antibodies in samples from 30 healthy volunteers who were orally infected with wild-type enterotoxigenic Escherichia coli. The response against these bacteria in serum, lavage, and lymphocyte supernatants (antibody-in-lymphocyte-supernatant, ALS) was compared with that in targeted parotid and sublingual/submandibular secretions. Strong correlation occurred between IgA antibody levels against the challenge bacteria in sublingual/submandibular secretions and in lavage (r=0.69, P<0.0001) and ALS (r=0.70, P<0.0001). In sublingual/submandibular secretions, 93% responded with more than a twofold increase in IgA antibodies against the challenge strain, whereas the corresponding response in parotid secretions was only 67% (P=0.039). With >twofold ALS as a reference, the sensitivity of a >twofold response for IgA in sublingual/submandibular secretion was 96%, whereas it was only 67% in the parotid fluid. To exclude that flow rate variations influenced the results, we used albumin as a marker. Our data suggested that IgA in sublingual/submandibular secretions, rather than whole saliva with its variable content of parotid fluid, is a preferential noninvasive proxy for intestinal immune induction.

Human IgG3 is decreased and IgG1, IgG2 and IgG4 are unchanged in molecular size by mild reduction and reoxidation without any major change in effector functions.

Purified proteins of the four human IgG subclasses were reduced under neutral conditions to break the interchain S-S bonds, followed by dialysis to allow reformation of S-S bonds (pr/o treatment). The IgG1, IgG2 and IgG4 proteins apparently reformed native molecules by pr/o treatment, while IgG3 formed molecules with significantly smaller size, as measured by HPLC gel filtration, compared to the autologous native proteins. The degree of shrinking of the pr/o IgG3 molecules varied and was most pronounced at low protein concn. In addition, the temp and the concn of reducing agent during the pr/o treatment had some influence on the molecular size. The effect is probably due to a conformational change of the 62 amino acid long hinge of IgG3. The effector activity of pr/o IgG2 and pr/o IgG3 was studied by employing chimeric, mouse V and human C regions, monoclonal antibodies with the same NIP-binding properties. Thus, the interaction between IgG and the complement system was unchanged both for pr/o IgG2 and pr/o IgG3, while the Fc-receptor-mediated antibody-dependent cellular cytotoxicity (ADCC) was depressed to the same degree for both pr/o IgG2 and pr/o IgG3. Conclusively, the alteration of the conformation of the IgG3 molecule by pr/o treatment had no major influence on its effector functions.

Antibody dependent cell-mediated cytotoxicity induced by chimeric mouse-human IgG subclasses and IgG3 antibodies with altered hinge region.

A matched set of chimeric mouse-human NP-antibodies were studied for the capacity to induce cell mediated cytotoxicity (ADCC) by normal peripheral blood NK/K cells. The target cells were sheep red blood cells (SRBC) sensitized with the haptens NP or NIP. All four IgG subclasses and several IgG3 variants with altered hinge were tested for ADCC activity. The hierarchy of the ADCC capacity among the subclasses was found to be IgG3 greater than IgG1 greater than IgG4 greater than IgG2. The superiority of IgG3 was only revealed at low effector cell:target cell ratio. The ADCC activity was for the most part unaltered by shortening the hinge region of IgG3 from 62 to 15 amino acids. Also, when the hinge region of IgG3 was mutated to become identical to that of IgG4, the ADCC activity was mainly unchanged. However, an IgG3 variant with deletion of all four hinge exons showed a depressed ADCC activity compared to the wild type. The IgG subclass pattern of complement-mediated lysis (CML) and ADCC is different and the capacity to induce CML and ADCC is changed differently by hinge region modification. Thus CML and ADCC have different structural requirements in the Fc region of IgG.

Human IgG3 can adopt the disulfide bond pattern characteristic for IgG1 without resembling it in complement mediated cell lysis.

In this paper we describe the construction of mouse-human IgG3 mutant antibodies resembling IgG1 in their disulfide bond pattern between the heavy and light chain (H-L) and between the two heavy chains (H-H). The effector functions of these mutant antibodies were compared to normal IgG3 and IgG1. Changing only the disulfide bond pattern between the heavy and light chains did not alter the ability to induce complement mediated cell lysis (CML), regardless of the amount of corresponding antigen that had been introduced to the surface of the target cells. However, alteration of the disulfide bond pattern between the two heavy chains had a large effect on CML due to shortening of the hinge from 62 to 15 amino acids. No difference between the mutants and normal antibodies in antibody-dependent cell-mediated cytotoxicity (ADCC) was observed. This suggests that IgG3 can adopt the H-L disulfide bond pattern of IgG1 without obtaining the CML activity characteristic for IgG1.

The use of a hapten-Fab conjugate to sensitize target cells for antibody-dependent complement-mediated lysis and antibody-dependent cell-mediated cytotoxicity.

NIP-conjugated Fab' fragments from a rabbit hyperimmunized with sheep red blood cells (SRBC) were used to link the hapten NIP to target cells for antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent complement-mediated lysis (ADCML). Target cells (SRBC) labelled with this NIP-Fab' complex were compared with SRBC directly haptenized with NIP in ADCC and ADCML assays using a NIP specific IgG1 chimeric antibody. Both methods yielded almost identical results. Using the NIP-FAb' conjugate identical target cell haptenization was readily achieved from experiment to experiment. Using conjugates of different NIP/Fab' ratios it should be possible to study how such changes influence antibody effector functions.

Unexpected interaction of some anti-TNP hybridoma antibodies with Superose HPLC gel filtration resins.

Five different hybridoma antibodies (the isotypes IgG1, IgG2a and IgG2b), reactive with TNP, showed increased elution times when gel filtered on a Superose-12 HPLC column corresponding to apparent molecular weights ranging from 54 kDa to 120 kDa compared to the normal of 150 kDa. One of the antibodies (C1901-B4) was studied in detail showing that the unusual gel filtration behaviour was localized to the Fab part of the molecule. SDS-PAGE analysis showed that the intact antibodies had normal molecular weights. Gel filtration on a Toyosoda G-3000SW HPLC column and Sephadex G-150 as well as Sepharose 6B generally showed normal elution times. These results support the hypothesis that the retardation on the Superose gel is probably due to aromatic interactions between amino acid residues supposedly exposed in the hypervariable region (i.e., the antibody combining site) of the antibody, and the gel matrix which is rich in ether O-atoms created during the manufacturing process. If this hypothesis is correct one might expect such interactions between Superose resins and antibodies of many different specificities in which aromatic amino acids are exposed in the combining sites.

Incidence of malignant melanoma of the skin in Norway, 1955-1989: associations with solar ultraviolet radiation, income and holidays abroad.

BACKGROUND: Norway has the highest incidence of melanoma in Europe. This study analyses geographical variations in melanoma incidence within Norway and their association with possible aetiological factors. METHODS: Data on melanoma incidence from the Norwegian Cancer Registry were used to calculate standardized incidence ratios (SIR) for the 19 counties in Norway for each 5-year period from 1955 to 1989. Multiple regression analysis was used to examine the associations between these SIR and local UVB levels, holidays abroad and income. Similar methods were also used to analyse changes in SIR between 1955-1969 and 1985-1989. RESULTS: There was a highly significant association between melanoma incidence and UVB in each of the time periods studied. Income showed a significant positive association in the 1960s and early 1970s but not later. Foreign holidays showed a significant positive association in the 1980s, but not earlier. Changes in melanoma SIR between 1955-1969 and 1985-1989 were significantly positively associated with holidays abroad and negatively with income levels. CONCLUSIONS: Melanoma incidence in Norway is closely related to local levels of UVB radiation independently of other factors suggesting that local exposures carry significant risk. Risks would probably increase if ozone depletion led to enhanced UVB flux (estimated as 1.6% rise in incidence for each 1% increase in UVB). By the end of the study period income was no longer a significant factor but holidays abroad had started to have a detectable effect on melanoma incidence.

Epitope analyses of pneumococcal surface protein A: a combination of two monoclonal antibodies detects 94% of clinical isolates.

Immunisation of BALB/c mice with seven heat-treated Norwegian clinical isolates of Streptococcus pneumoniae of different serotypes elicited mainly monoclonal antibodies (mAbs) to pneumococcal surface protein A (PspA). It was remarkable that the fusions resulted only in a few mAbs directed against other protein antigens. Dot blot analysis with 16 mAbs using clinical isolates representing 23 different capsular types and the uncapsulated reference strain R36A showed that some of the mAbs bound to PspA epitopes expressed by a low number of strains whereas others bound to broadly distributed epitopes. On the basis of their reactivities, seven of these mAbs could be divided into two groups recognising different subsets of pneumococci. The three mAbs in the narrow reacting group bound to epitopes found in 21-25% of the strains whereas the four mAbs in the broad reacting group detected more than 57% of the analysed strains. The epitopes for these seven antibodies were surface exposed on live exponential phase grown pneumococci as shown by flow cytometry. The finding that a combination of mAb 180,C-1 (IgG2a) from the first group and mAb 170,E-11 (IgG2a) from the second group detected 94% of the examined strains is interesting because PspA has been reported by others to be a serological highly variable protein.

Streptococcus pneumoniae heat shock protein 70 does not induce human antibody responses during infection.

Mouse monoclonal antibodies (mAbs) were developed against Streptococcus pneumoniae in search for potential common pneumococcal proteins as vaccine antigens. mAb 230,B-9 (IgG1) reacted by immunoblotting with a 70-kDa protein which was isolated by immunoaffinity chromatography and subsequent preparative electrophoresis. N-terminal amino acid sequencing showed homology to that of heat shock protein 70 (hsp70). The hsp70 epitope reactive with mAb 230,B-9 was found in all the pneumococci examined as well as in other streptococci and enterococci. The epitope was not expressed in several other examined Gram-positive or -negative bacteria. Pneumococcal hsp70 has by other investigators been proposed to be a vaccine candidate. Binding experiments using flow cytometry showed that the epitope was not surface-exposed on live exponential phase grown S. pneumoniae. Human patient sera did not react with affinity-purified pneumococcal hsp70. Therefore the pneumococcal hsp70 does not seem to be of special interest in a vaccine formulation. The human sera contained antibodies to high molecular proteins co-purified with hsp70. Some of these proteins could be the pneumococcal surface protein A.

Ischaemic heart disease mortality among men in Norway: reversal of urban-rural difference between 1966 and 1989.

OBJECTIVE: This study aimed to examine regional urban-rural differences in mortality from ischaemic heart disease, including sudden death of unknown cause (IHD/SUD) in Norway from 1966-89, for men and women aged 30-69 years. DESIGN: Analysis was based on vital statistics. Regional mortality rates were obtained by aggregating the 443 municipalities in Norway into urban, rural, and intermediate municipalities. SETTINGS AND SUBJECTS: Norway. RESULTS: In 1966-70 the age adjusted IHD/SUD mortality in the age group 30-69 years was higher in urban than in rural areas; for men by 31% (95% CI 27%, 36%) and for women by 28% (95% CI 19%, 36%). In 1986-89 the IHD/SUD mortality for men showed a reversed urban-rural gradient: it was 8% (95% CI 2%, 13%) higher in rural than in urban areas. The mortality rates for women were equal for both these aggregates. For men the results indicate that IHD/SUD mortality peaked first in urban municipalities and then, but at a lower level, in rural areas. For women there was a substantial decline in IHD/SUD mortality between 1966 and 1989, but an actual peak could not be demonstrated in any of the three aggregates during the period. The decline in IHD/SUD mortality among women was steepest in urban municipalities and least noticeable in rural municipalities, but the decline tapered off towards the end of the study period. CONCLUSION: The results confirm a phase-shifted peak in IHD/SUD mortality, which began in towns and ended in rural areas, and provides clues to the main underlying factors in the IHD epidemic at the population level.

Regionalizing mortality data: ischaemic heart disease in Norway.

The way regions are delimited has a bearing on the geographical patterns and time trends which emerge from cause specific mortality analysis. Whenever possible, alternative regionalizations should be used to explore the full information potential of the data. For statistical reasons, the size of the regional units (populations-at-risk) should be selected according to the frequency of the cause of death, number of years in the time period, etc. A geographical mortality information system for Norway, based on individual death records and with quick and flexible retrieval options is described. As a demonstration, geographical time trends in ischaemic heart disease from 1970 to 1985 are studied, using different schemes of regionalization. A clear tendency towards regional convergence appears in the rural-urban dimension, but there is no convergence between the five subnational regions of the country. There is no evidence that counties which have received heart disease intervention projects fare any better than those which have not, but here a more thorough analysis is recommended. Within the intervention counties, there are large variations both in mortality levels and trends.

The diffusion of cardiovascular disease in the Norwegian farming community: a combination of morbidity and mortality data.

This article analyses an observed increase in cardiovascular morbidity among male farmers in Norway during the last decade in the light of the traditionally low mortality in farmers. Three hypotheses to explain the increases in CVD morbidity are tested, of which one, stating that there is a time lag in the spread of risk factors, proves to be most fruitful. Mortality data for agricultural communities show no increase in overall CVD rates, but when age-specific rates are analysed, an increase in the younger age groups emerges, especially for ischaemic heart disease. If this process continues, farmers and farming areas may change from low to high mortality, relatively speaking. It is argued that this change is due to a time lag in two waves, first an increase in risk factors such as smoking, more fatty diets and less physically demanding work, then improved lifestyles due to a better perception of risk factors. Both waves may be affecting rural areas later than the urban centres. Knowledge of such geographical and socio-economic diffusion processes is important in the planning and implementation of prevention programmes.

Gender, geography and socio-economic status in the diffusion of malignant melanoma risk.

Malignant melanoma is the cancer that has shown the fastest increase in incidence in most white populations in recent decades. This paper studies the diffusion of the disease for males and females, geographical areas and socio-economic groups. Incidence data from the Norwegian Cancer Registry covering the period 1955-1989 make it possible to establish birth cohorts covering a time span from the late 1880s to the late 1950s. For Norway as a whole the increase in incidence was apparent between cohorts born in the 1880s and the 1890s for both sexes. The epidemic was first observable in the most central, south-eastern part of the country, with the onset reaching the most peripheral Northern Norway from the cohort born in the late 1910s. The onset also came later in rural than urban Norway. There are indications that the epidemic started in the more affluent part of the population, followed by an equalization. The increase in melanoma could be a result of more active outdoor recreation from the end of the last century. In cohorts born from the 1930s onwards a slowing down in the increase can be observed. International comparisons show that in most countries the incidence pattern in the first decades of the epidemic was dominated by males; later on the incidence became higher for females. Many cancer registries in the U.S.A. and Oceania now show a downward trend in melanoma incidence for males and females aged 15-29 years. A similar development has not been observed in Europe and Canada up to 1983-1987.

Antibody-induced opsonophagocytosis of serogroup B meningococci measured by flow cytometry.

Antibodies can protect against meningococcal infection by at least two mechanisms: complement-dependent serum bactericidal activity (SBA), and/ or opsonophagocytosis (OP), leading to destruction of the bacteria (1-4). Regarding group C meningococci, there seems to be a correlation between SBA activity and protection (1), whereas OP has been less well-studied. This function may have a supplementary or even major role in protection against group B meningococci.

T-cell responses against meningococcal antigens.

T-cells recognize protein antigens as short peptide fragments (8-20 amino acids) bound to major histocompatibility complex (MHC) molecules on the surface of antigen-presenting cells (APCs). A prerequisite for antigen-specific T-cell activation is antigen uptake, enzymatic degradation, and recycling of MHC-peptide complexes to the surface of APCs. Whereas CD8+ T cells recognize endogenously derived antigen (virus and other intracellular pathogens) bound to MHC class I molecules, CD4+ T cells recognize exogenously derived antigen in complex with MHC class II molecules. Hence, extracellular bacteria, such as meningococci during invasive disease, will be presented to CD4+ T cells in the context of MHC class II molecules, after uptake and processing by professional APCs like B cells, macrophages, or dendritic cells. Antigen-specific CD4+ T cells can be classified as Th1 or Th2 subpopulations on the basis of different cytokine production and effector functions (1). Intracellular microbes often induce Th1-dominated responses, whereas extracellular pathogens and parasites typically trigger Th2 responses. Th1 cells produce mainly interleukin (IL)-2, interferon (IFN)-γ, and tumor necrosis factor (TNF)-ß, which represent important inducers of the cell-mediated immune responses. The principal Th1 cytokine IFN-γ activates macrophages by enhancing their ability to phagocytize and destroy microbes by intracellular bactericidal mechanisms. In contrast, Th2 cells produce IL-4, IL-5, IL-6, and IL-13, which are important factors for inducing and regulating B-cell responses (1).

Mechanism for uptake of silica particles by monocytic U937 cells.

We examined the mechanism for uptake by monocytic cells of particles found in the atmosphere of some industrial work places. As a model system, irregular crystalline silica particles (SPs), sphere-like cryptocrystalline microsilica particles (MPs) and carbon particles (CPs) were exposed to pro-monocytic U937 cells. Plasma-treated SP and MP, but not CP, activated the alternative complement pathway, but bound little C3b. However, all particles adsorbed serum IgG, IgA and IgM unspecifically. Phenotyping of U937 cells for complement receptors (CRs) and Fcgamma receptors (FcgammaRs) showed that interferon gamma (INFgamma) increased expression of FcgammaRI, CR3 (CD11b/CD18) and CR4 (CD11c/CD18) and that phorbol-12-myristate-13-acetate (PMA) increased expression of CR4. Scanning electron microscopy (SEM) demonstrated higher phagocytosis of plasma-treated SP than native SP by both PMA- and INFgamma-stimulated, but not unstimulated, cells. MP and CP could not be distinguished from cellular structures. Inhibition experiments in SEM revealed uptake of heparin-plasma-treated SP via FcgammaRI on INFgamma-stimulated U937 cells, but could not exclude possible participation of CR3. The results indicate that plasma-treated SPs bind Ig and are internalized by differentiated monocytic cells via FcgammaRI, which is known to trigger cellular production of toxic oxygen species that may induce pulmonary inflammation in vivo.

Preclinical immunogenicity study of trivalent meningococcal AWX-OMV vaccines for the African meningitis belt.

In the recent decade, epidemic meningitis in the African meningitis belt has mostly been caused by Neisseria meningitidis of serogroups A, W and X (MenA, MenW and MenX, respectively). There is at present no licensed vaccine available to prevent MenX meningococcal disease. To explore a trivalent MenAWX vaccine concept, we have studied the immunogenicity in mice of MenX outer membrane vesicles (X-OMV) or MenX polysaccharide (X-PS) when combined with a bivalent A-OMV and W-OMV (AW-OMV) vaccine previously shown to be highly immunogenic in mice. The vaccine antigens were produced from three representative wild type strains of MenA (ST-7), MenW (ST-11) and MenX (ST-751) isolated from patients in the African meningitis belt. Groups of mice were immunized with two doses of X-OMV or X-PS combined with the AW-OMV vaccine or as individual components. All vaccine preparations were adsorbed to Al(OH)3. Sera from immunized mice were tested by ELISA and immunoblotting. Functional antibody responses were measured as serum bactericidal activity (SBA) and opsonophagocytic activity (OPA). Immunization of mice with X-OMV, alone or in combination with AW-OMV induced high levels of anti-X OMV IgG. Moreover, X-OMV alone or in combination with the AW-OMV vaccine induced high SBA and OPA titers against the MenX target strain. X-PS alone was not immunogenic in mice; however, addition of the AW-OMV vaccine to X-PS increased the immunogenicity of X-PS. Both AWX vaccine formulations induced high levels of IgG against A- and W-OMV and high SBA titers against the MenA and MenW vaccine strains. These results suggest that a trivalent AWX vaccine, either as a combination of OMV or OMV with X-PS, could potentially prevent the majority of meningococcal disease in the meningitis belt.

Preclinical immunogenicity and functional activity studies of an A+W meningococcal outer membrane vesicle (OMV) vaccine and comparisons with existing meningococcal conjugate- and polysaccharide vaccines.

Meningococci of serogroups A and W (MenA and MenW) are the main causes of epidemic bacterial meningitis outbreaks in sub-Saharan Africa. In this study we prepared a detergent extracted outer membrane vesicle (dOMV) vaccine from representative African MenA and MenW strains, and compared the immunogenicity of this vaccine with existing meningococcal conjugate and polysaccharide (PS) vaccines in mice. NMRI mice were immunized with preclinical batches of the A+W dOMV vaccine, or with commercially available vaccines; a MenA conjugate vaccine (MenAfriVac(®), Serum Institute of India), ACYW conjugate vaccine (Menveo(®), Novartis) or ACYW PS vaccine (Mencevax(®), GlaxoSmithKline). The mice received 2 doses of 1/10 or 1/50 of a human dose with a three week interval. Immune responses were tested in ELISA, serum bactericidal activity (SBA) and opsonophagocytic activity (OPA) assays. High levels of IgG antibodies against both A and W dOMV were detected in mice receiving the A+W dOMV vaccine. High SBA titers against both MenA and MenW vaccine strains were detected after only one dose of the A+W dOMV vaccine, and the titers were further increased after the second dose. The SBA and OPA titers in mice immunized with dOMV vaccine were significantly higher than in mice immunized with the ACYW-conjugate vaccine or the PS vaccine. Furthermore, the A+W dOMV vaccine was shown to induce SBA and OPA titers against MenA of the same magnitude as the titers induced by the A-conjugate vaccine. In conclusion, the A+W dOMV vaccine induced high levels of functional antibodies to both MenA and MenW strains, levels that were shown to be higher or equal to the levels induced by licensed meningococcal vaccines. Thus, an A+W dOMV vaccine could potentially serve as an alternative or a supplement to existing conjugate and PS vaccines in the African meningitis belt.

Investigation of different group A immunoassays following one dose of meningococcal group A conjugate vaccine or A/C polysaccharide vaccine in adults.

A double-blind, randomized, controlled phase I study to assess the safety, immunogenicity, and antibody persistence of a new group A conjugate vaccine (PsA-TT) in volunteers aged 18 to 35 years was previously performed. Subjects received one dose of either the PsA-TT conjugate vaccine, meningococcal A/C polysaccharide vaccine (PsA/C), or tetanus toxoid vaccine. The conjugate vaccine was shown to be safe and immunogenic as demonstrated by a standardized group A-specific immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and by a serum bactericidal antibody (SBA) assay using rabbit complement (rSBA). This report details further analysis of the sera using four additional immunologic assays to investigate the relationship between the different immunoassays. The immunoassays used were an SBA assay that used human complement (hSBA), a group A-specific IgG multiplexed bead assay, and two opsonophagocytic antibody (OPA) assays which used two different methodologies. For each vaccine group, geometric mean concentrations or geometric mean titers were determined for all assays before and 4, 24, and 48 weeks after vaccination. Pearson's correlation coefficients were used to assess the relationship between the six assays using data from all available visits. An excellent correlation was observed between the group A-specific IgG concentrations obtained by ELISA and those obtained by the multiplexed bead assay. hSBA and rSBA titers correlated moderately, although proportions of subjects with putatively protective titers and those demonstrating a > or = 4-fold rise were similar. The two OPA methods correlated weakly and achieved only a low correlation with the other immunoassays. The correlation between hSBA and group A-specific IgG was higher for the PsA-TT group than for the PsA/C group.