Burden of non-communicable diseases and the achievement of the sustainable development goals in 2030 in Mercosur countries.

OBJECTIVES: The aims of this article were to analyse the burden of NCDs and their RFs in the Mercosur countries between 1990 and 2019 and to project mortality trends for 2030. STUDY DESIGN: Epidemiological study of time series. METHODS: The present study used data from the Global Burden of Disease study. The absolute number of deaths, mortality rates, disability-adjusted life years, years of life lost, years lived with disability and the burden of premature mortality by NCD attributable to the RFs were evaluated. Projections were made up to 2030. Age-standardised rates were used to draw comparisons by years and by countries. The analysis was conducted using the RStudio software. RESULTS: Between 1990 and 2019, a decrease was found in the premature mortality rates caused by NCDs in all the countries, except for Paraguay, which remained stable. When analysing premature mortality rates due to NCDs up to 2030, it was predicted that none of the countries would achieve the sustainable development goal of a one-third reduction in premature mortality by NCDs. Regarding the impacts of the RFs for NCDs, smoking, dietary risks, high blood pressure (BP) and high body mass index (BMI) were the main risks attributable to premature deaths due to NCDs. CONCLUSIONS: The results showed that mortality rates are declining in Mercosur countries; however, none of the countries are predicted to achieve the sustainable development goal of a one-third reduction in mortality due to NCDs by 2030. In addition to access to adequate treatment, progress is required in public regulation actions to reduce RFs, such as smoking, dietary risks, high BP and high BMI.

Chronic Aerobic Training at Different Volumes in the Modulation of Macrophage Function and in vivo Infection of BALB/c Mice by Leishmania major.

Physical inactivity is one of the main causes of chronic diseases; however, strenuous exercise can induce immunosuppression. Several studies suggest that moderate amounts of exercise lead to a Th1 response, favoring the resolution of infections caused by intracellular microorganisms, while high volumes of exercise tend to direct the response to Th2, favoring infection by them. Leishmaniasis is a parasitic disease promoted by parasites of the Leishmania genus, with clinical manifestations that vary according to the species of the parasite and the immune response of the host. The experimental Leishmania major-BALB/C mouse model provides a good model for the resistance (Th1 response) or susceptibility (Th2 response) that determines the progression of this infection. The aim of this study was to evaluate the effect of aerobic training at different volumes on modulation of in vitro macrophage infection by L. major, as well as to assess the effect of high volume (HV) aerobic training on the development of L. major in vivo in BALB/c mice. Uninfected animals were submitted to various exercise volumes: none (SED), light (LV), moderate (MV), high (HV), very high (VHV), and tapering (TAP). The macrophages of these animals were infected by L. major and the LV and MV groups showed a decrease in the infection factor, while the VHV showed an increase in the infection factor, when treated with LPS. The cytokine concentration pattern measured in the supernatants of these macrophages suggested a predominant Th1 response profile in the LV and MV groups, while the Th2 profile predominated in the VHV and TAP groups. Groups of BALB/C mice infected with L. major were subjected to high volume (iHV) or non-periodized high volume (iNPHV) exercise or kept sedentary (iSED). The exercised animals suffered a significant increase in injuries caused by the parasites. The animals in the group submitted to high volume exercise (iHV) showed visceralization of the infection. These data strongly suggest that a very high volume of aerobic training increased the susceptibility of BALB/C mice to L. major infection, while moderate distribution of training loads promoted immunological balance, better controlling the infection by this parasite.

Protective immunity against malaria after vaccination.

A good understanding of the immunological correlates of protective immunity is an important requirement for the development of effective vaccines against malaria. However, this concern has received little attention even in the face of two decades of intensive vaccine research. Here, we review the immune response to blood-stage malaria, with a particular focus on the type of vaccine most likely to induce the kind of response required to give strong protection against infection.

Effect of calcium hydroxide and endodontic irrigants on fibre post bond strength to root canal dentine.

AIM: To analyse the effect of calcium hydroxide paste, endodontic irrigants and time of application on the bond strength of fibre posts to root canal dentine. METHODOLOGY: Seventy bovine incisors were divided into 7 groups according to removal of calcium hydroxide and distilled water (CHW) (immediate - I; 21 days - 21 days; 6 months - 6 months) and endodontic irrigant (1% sodium hypochlorite - SH; 1% sodium hypochlorite + 17% EDTA - SHE). Fibre posts were cemented (RelyX U100), after which the roots were serially sectioned and submitted to a micro-push-out test. Data were analysed using two-way anova followed by the Tukey's and the Dunnett's tests (α = 0.05). RESULTS: The CHW groups were not significantly different from the control group in 13 of the 18 associated factors (P > 0.05). There were significant reductions in bond strength in the cervical (P = 0.0216) and middle (P = 0.0017) thirds of the root at 6 months in groups irrigated with SH. Irrigation with SHE reduced the bond strength significantly in the middle (P = 0.0488) and apical (P = 0.0252) thirds of the roots in the immediate groups and in the middle third (P = 0.0287) in the 21-day group. Bond strength was greater in the cervical than in the apical thirds of all immediate and 21-day groups (P < 0.05). CONCLUSIONS: Bond strength of groups that received CH paste was similar to that found in the control group in 13 of the 18 associated factors. EDTA and SH reduced bond strength in specimens in the immediate (middle and apical thirds) and 21-day (middle third) groups. There was a significant reduction in bond strength in the groups irrigated with SH and tested at 6 months (cervical and middle thirds). There was a predominance of adhesive failures between resin cement and dentine in all groups.

Endogenous galectin-3 controls experimental malaria in a species-specific manner.

Galectins are evolutionarily conserved glycan-binding proteins with pleiotropic roles in innate and adaptive immune responses. Galectin-3 has been implicated in several immunological processes as well as in pathogen recognition through specific binding to glycosylated receptors on the surface of host cells or microorganisms. In spite of considerable evidence supporting a role for galectin-3 in host-pathogen interactions, the relevance of this lectin in the regulation of the host defence mechanisms in vivo is poorly understood. In this study, we analysed the impact of galectin-3 deficiency during infection with three distinct species of rodent malaria parasites, Plasmodium yoelii 17XNL, Plasmodium berghei ANKA and Plasmodium chabaudi AS. We found that galectin-3 deficiency showed a marginal effect on the course of parasitaemia during P. chabaudi infection, but did not alter the course of parasitaemia during P. berghei infection. However, lack of galectin-3 significantly reduced P. yoelii parasitaemia. This reduced parasitaemia in Lgals3(-/-) mice was consistent with higher titres of anti-P. yoelii MSP1(19) IgG2b isotype antibodies when compared with their wild-type counterparts. Our results reflect the complexity and singularity of host-pathogen interactions, indicating a species-specific role of endogenous galectin-3 in the control of parasite infections and the modulation of antibody responses.

Effect of ozone application on the resin-dentin microtensile bond strength.

When ozone is used during caries treatment, bond strength can be compromised by the release of oxygen. The use of antioxidant agents neutralizes the free oxygen. The aim of this study was to evaluate the effects of ozone and sodium ascorbate on resin-dentin microtensile bond strength (µTBS). Forty human third molars were divided into four groups: Group 1, not treated with ozone; Group 2, ozone application followed by acid etching; Group 3, acid etching followed by ozone application; and Group 4, ozone and application of sodium ascorbate. Bonded beams (1.0 mm(2)) were tested under tension (0.5 mm min(-1)). The µTBS values were analyzed using one-way analysis of variance (ANOVA) and the Tukey test (p=0.05). All beams that fractured were analyzed under stereomicroscopy (40×). Group 1 had significantly higher µTBS values than Group 2 or 3. The µTBS values of Groups 1 and 4 were similar and higher than those of Group 2. The use of ozone in Group 2 resulted in lower values of µTBS in all conditions evaluated. The predominant failure mode was adhesive. The application of ozone decreased the µTBS of the dentin-composite resin interface. These values were reversed when compared with Groups 1 and 2 when sodium ascorbate was used.

In vitro inhibition of oral streptococci binding to the acquired pellicle by algal lectins.

AIMS: The initial colonization of the tooth by streptococci involves their attachment to adsorbed components of the acquired pellicle. Avoiding this adhesion may be successful in preventing caries at early stages. Salivary mucins are glycoproteins that when absorbed onto hydroxyapatite may provide binding sites for certain bacteria. Algal lectins may be especially interesting for oral antiadhesion trials because of their great stability and high specificity for mucins. This work aimed to evaluate the potential of two algal lectins to inhibit the adherence of five streptococci species to the acquired pellicle in vitro. METHODS AND RESULTS: The lectins used were extracted from Bryothamnion triquetrum (BTL) and Bryothamnion seaforthii (BSL). Fluorescence microscopy was applied to visualize the ability of fluorescein isothiocyanate-labelled lectins to attach to the pellicle and revealed a similar capability for both lectins. Streptococcal adherence assays were performed using saliva-coated microtitre plates. BSL inhibited more than 75% of Streptococcus sanguis, Streptococcus mitis, Streptococcus sobrinus and Streptococcus mutans adherence, achieving 92% to the latter. BTL only obtained statistically significant results on S. mitis and S. sobrinus, whose adherence was decreased by 32.5% and 54.4%, respectively. CONCLUSION: Algal lectins are able to inhibit streptococcal adherence. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results support the proposed application of lectins in antiadhesion therapeutics.

Monitoring the trihalomethanes present in water after treatment with chlorine under laboratory condition.

In this work assays involving chlorinated water samples, which were previous spiked with humic substances or algae blue green and following the production of the THMs for 30 days is described. To implement the assays, five portions of 1,000 ml of water were stored in glass bottles. The water samples were treated with solutions containing 2, 3, 4 and 5 mg l(-1) chlorine. The samples aliquots (60 ml) were transferred into the glass vials, 10 ml were removed to have a headspace and 100 microl of the 10 mg l(-1) pentafluortoluene bromide solution was added to each vial. The extraction step was performed by adding 10 g of Na(2)SO(4) followed by 5 ml of n-pentane. The vials were stopped with a TFE-faced septum and sealed with aluminum caps. The generated THMs were determined by gas chromatography with electron capture detector using reference solutions with concentration ranging from 8 to 120 microg l(-1) THMs. Three assays were monitored during 30 days and chloroform was the predominant compound found in the water samples, while other species of THMs were not detected. The results showed that when the chlorine concentration was increased in water samples containing algae the concentration of THM varied randomly. Nevertheless, in water samples containing humic substances the increase of the THM concentration presented a relationship with the chlorine concentration. It was also observed that chloroform concentration increased with the elapsed time up to one and six days to water samples spiked with humic substances and algae blue green, respectively and decreased along 30 days. By other hand, assays performed using water samples containing decanted algae material showed that THM was not generated by the chlorine addition.

Inhibition of NTPDase, 5'-nucleotidase, Na+/K+-ATPase and acetylcholinesterase activities by subchronic treatment with Casearia sylvestris.

The aqueous extract of Casearia sylvestris was tested in cortical membrane preparations. C. sylvestris was obtained commercially from two different sources, designated as Sample A and Sample B. The enzymes studied in this work were NTPDase-like, 5'-Nucleotidase, Na(+)/K(+)-ATPase and acetylcholinesterase (AChE). Adult rats received aqueous extracts from C. sylvestris in a dose of 20mg/kg body wt. daily for a 75-day-period, by oral administration (gavage). Our study showed that this treatment caused an inhibition of NTPDase-like activity with both, ATP (19.41% with Sample A and 25.03% with Sample B) and ADP (41.57% with Sample A and 31.20% with Sample B) as substrates. This treatment also caused an inhibition of 5'-nucleotidase activity (28.34% with Sample A and 31.46% with Sample B) and Na(+)/K(+)-ATPase (25.08% with Sample A and 24.81% with Sample B). The rate of acetylcholine degradation was reduced, as shown by the inhibition of AChE (31.65% and 26.74%, Samples A and B, respectively). These results suggest that extracts of C. sylvestris can cause neurochemical alterations in the purinergic and cholinergic systems of the central nervous system.

Glycosylphosphatidylinositols in malaria pathogenesis and immunity: potential for therapeutic inhibition and vaccination.

Glycosylphosphatidylinositols (GPIs) are found in the outer cell membranes of all eukaryotes. GPIs anchor a diverse range of proteins to the surface of Plasmodium falciparum, but may also exist free of protein attachment. In vitro and in vivo studies have established GPIs as likely candidate toxins in malaria, consistent with the prevailing paradigm that attributes induction of inflammatory cytokines, fever and other pathology to parasite toxins released when schizonts rupture. Although evolutionarily conserved, sufficient structural differences appear to exist that impart upon plasmodial GPIs the ability to activate second messengers in mammalian cells and elicit immune responses. In populations exposed to P. falciparum, the antibody response to purified GPIs is characterised by a predominance of immunoglobulin (Ig)G over IgM and an increase in the prevalence, level and persistence of responses with increasing age. It remains unclear, however, if these antibodies or other cellular responses to GPIs mediate anti-toxic immunity in humans; anti-toxic immunity may comprise either reduction in the severity of disease or maintenance of the malaria-tolerant state (i.e. persistent asymptomatic parasitaemia). P. falciparum GPIs are potentially amenable to specific therapeutic inhibition and vaccination; more needs to be known about their dual roles in malaria pathogenesis and protection for these strategies to succeed.

Improvement of glucose homeostasis in obese diabetic db/db mice given Plasmodium yoelii glycosylphosphatidylinositols.

We have previously reported that infection with Plasmodium yoelii, Plasmodium chabaudi, or injection of extracts from malaria-parasitized red blood cells induces hypoglycemia in normal mice and normalizes the hyperglycemia in streptozotocin (STZ)-diabetic mice. P yoelii glycosylphosphatidylinositols (GPIs) were extracted in chloroform:methanol:water (CMW) (10:10:3), purified by high-performance thin layer chromatography (HPTLC) and tested for their insulin-mimetic activities. The effects of P yoelii GPIs on blood glucose were investigated in insulin-resistant C57BL/ks-db/db diabetic mice. A single intravenous injection of GPIs (9 and 30 nmol/mouse) induced a significant dose-related decrease in blood glucose (P < .001), but insignificantly increased plasma insulin concentrations. A single oral dose of 2.7 micromol GPIs per db/db mouse significantly lowered blood glucose (P < .01). P yoelii GPIs in vitro (0.062 to 1 micromol/L) significantly stimulated lipogenesis in rat adipocytes in a dose-dependent manner both in the presence and absence of 10(-8) mol/L insulin (P < .01). P yoelii GPIs stimulated pyruvate dehydrogenase phosphatase (PDH-Pase) and inhibited both cyclic adenosine monophosphate (cAMP)-dependent protein kinase A and glucose-6-phosphatase (G6Pase). P yoelii GPIs had no effect on the activity of the gluconeogenic enzymes fructose-1,6-bisphosphatase (FBPase) and phosphoenolpyruvate carboxykinase (PEPCK). This is the first report of the hypoglycemic effect of P yoelii GPIs in murine models of type 2 diabetes. In conclusion, P yoelii GPIs demonstrated acute antidiabetic effects in db/db mice and in vitro. We suggest that P yoelii GPIs, when fully characterized, may provide structural information for the synthesis of new drugs for the management of diabetes.

A prospective open-label study to evaluate the effects of the oral contraceptive Harmonet (gestodene75/EE20) on body fat.

This open-label study evaluated the effects on body fat of the use of a low-dose oral contraceptive (gestodene75/EE20) in a group of 61 women (OC-U group) as compared to a nonuser group (OC-N group) of 51 women who did not receive an oral contraceptive. Weight, body mass index (BMI), waist-over-hip ratio and body composition data, obtained by bioelectrical impedance [percentages of body fat (%FAT), water (%TBW) and lean mass (%FFM)], were assessed before and after six treatment cycles. Baseline OC-U group weight, BMI, %FAT, %TBW and %FFM did not differ from the OC-N group, either at baseline or at the end of the study, and did not significantly change within each group during the study. Also, there was no modification of fat distribution in either group. Among women in the OC-U group, there was a slight increase in total cholesterol levels and a trend towards higher triglycerides levels. No changes were detected in blood pressure. In conclusion, this low-dose oral contraceptive did not affect weight or body composition. Thus, our data suggest that gestodene75/EE20 represents an appropriate OC choice and may enhance compliance of women who mistakenly believe that the use of oral contraceptives always leads to weight gain.

Effect of gamma radiation on dentin bond strength and morphology.

Sterilization by gamma radiation is a method often used for bone and extracted teeth banking. The bond strength of human dentin submitted to gamma rays has not been reported. Therefore, the aim of this study was to assess the effect of gamma radiation on dentin shear bond strength and morphology. The roots were removed from extracted human bicuspids and their crowns divided into two groups: an untreated control and crowns submitted to gamma radiation sterilization. The crowns were mounted in epoxy resin and the buccal enamel removed exposing the subjacent dentin. SBMPPlus adhesive system was applied to a 3-mm diameter area after 15 s of 35% phosphoric acid etching. The samples were mounted in composite resin cylinders and stored in distilled water at 37 degrees C for 24 h until the shear test. Dental fragments from both groups were prepared for SEM analysis. There was no statistically significant difference between the results of the shear test for the two groups according to the Tukey test (p > 0.05). Scanning electron micrographs also did not show alterations. These results indicate that gamma radiation neither affected the shear bond strength of SBMPPlus nor altered the dentin surface morphology.

Long-term prognosis of cirrhosis after spontaneous bacterial peritonitis treated with ceftriaxone.

Spontaneous bacterial peritonitis (SBP) is a frequent infection in cirrhotic patients with ascites, with a poor prognosis. The aims of this study were to determine the long-term survival of cirrhotic patients with SBP treated with ceftriaxone and to identify predictive factors related to survival. We studied 47 first episodes of SBP treated with ceftriaxone with a mean follow-up of 272 days. Nineteen variables were recorded to evaluate their relation to survival. The most frequent organism that caused SBP was Escherichia coli (40%). Spontaneous bacterial peritonitis resolution was achieved in 67% of patients. After resolution, SBP recurrence was observed in 44% of patients. The cumulative probability of survival was 68.1% at 1 month and 30.8% at 6 months. After uni-and multivariate analyses of all cases, SBP resolution ( p = 0.0001) and international normalized ratio (INR) ( p = 0.0057) were found to be related to survival. Another analysis performed after SBP resolution and SBP recurrence showed that ascitic fluid-positive culture ( p = 0.0344) and INR ( p = 0.0218) had statistical significance as variables predictive of long-term survival. We conclude that the survival of cirrhotic patients is very short after the first episode of SBP, a fact probably related to advanced liver disease, as liver dysfunction (INR) is the most important factor related to long-term patient survival.

Reversal of type 2 diabetes in mice by products of malaria parasites. II. Role of inositol phosphoglycans (IPGs).

We have previously shown that infection with Plasmodium yoelii malaria or injection of extracts from malaria-parasitized red cells induces hypoglycemia in normal mice and normalizes the hyperglycemia in mice made moderately diabetic with streptozotocin. Inositol phosphoglycans (IPGs) are released outside cells by hydrolysis of membrane-bound glycosylphosphatidylinositols (GPIs), and act as second messengers mediating insulin action. The C57BL/Ks-db/db and C57BL/6J-ob/ob mice offer good models for studies on human obesity and Type 2 diabetes. In the present study, we show that a single iv injection of IPG-A or IPG-P extracted from P. yoelii significantly (P < 0.02) lowers the blood glucose in STZ-diabetic, db/db, and in ob/ob mice for at least 4--6 h. Using rat white adipocytes, IPG-P increased lipogenesis by 20--30% in the presence and absence of maximal concentrations of insulin (10(-8) M) (P < 0.01) and stimulated pyruvate dehydrogenase (PDH) phosphatase in a dose-related manner. Both IPG-A and IPG-P inhibited c-AMP-dependent protein kinase (PKA) in a dose-related manner. Compositional analysis of IPGs after 24 h hydrolysis revealed the presence of myo-inositol, phosphorus, galactosamine, glucosamine, and glucose in both IPG-A and IPG-P. However, hydrolysis of IPGs for 4 h highlighted differences between IPG-A and IPG-P. There are some functional similarities between P. yoelii IPGs and those previously described for mammalian liver. However, this is the first report of the hypoglycemic effect of IPGs in murine models of Type 2 diabetes. We suggest that IPGs isolated from P. yoelii, when fully characterized, may provide structural information for the synthesis of new drugs for the management of diabetes mellitus.

Gingival erosive lichen planus: case report.

A case of gingival erosive lichen planus is presented with special emphasis on its clinical and microscopic characteristics. The differential diagnosis and the controversy associated with the malignant potential of oral lichen planus is also discussed.

Early nonspecific immune responses and immunity to blood-stage nonlethal Plasmodium yoelii malaria.

The early role of natural killer cells and gamma delta T cells in the development of protective immunity to the blood stage of nonlethal Plasmodium yoelii infection was studied. Splenic cytokine levels were measured 24 h after infection of natural killer cell-depleted immunodeficient and littermate mice or transiently T-cell-depleted normal mice. Splenic gamma interferon levels were significantly increased above background in immunodeficient and littermate mice 24 h after infection. Depletion of natural killer cells resulted in markedly depressed gamma interferon levels and poor control of parasitemia, particularly in severe combined immunodeficient mice. In the littermates, gamma interferon levels were partially reduced, but parasitemias were resolved normally. However, in athymic mice, natural killer cell depletion had no effect on gamma interferon production. Levels of tumor necrosis factor alpha were increased in all animals 24 h after infection, and responses were not affected by natural killer cell depletion. However, in T-cell-depleted animals, both gamma interferon and tumor necrosis factor alpha levels were decreased 24 h after infection, and depleted mice were unable to control their parasitemia. These results suggest that the early production of both cytokines is important in the early control of parasitemia and that both natural killer and gamma delta T cells contribute equally towards their production. The data also suggest that the subsequent resolution of infection requires early production of gamma interferon, which might act by switching on the appropriate T-helper-cell subsets and other essential parasitotoxic effector mechanisms.

Immunopathology of cerebral malaria: morphological evidence of parasite sequestration in murine brain microvasculature.

A murine model that closely resembles human cerebral malaria is presented, in which characteristic features of parasite sequestration and inflammation in the brain are clearly demonstrable. "Young" (BALB/c x C57BL/6)F(1) mice infected with Plasmodium berghei (ANKA) developed typical neurological symptoms 7 to 8 days later and then died, although their parasitemias were below 20%. Older animals were less susceptible. Immunohistopathology and ultrastructure demonstrated that neurological symptoms were associated with sequestration of both parasitized erythrocytes and leukocytes and with clogging and rupture of vessels in both cerebral and cerebellar regions. Increases in tumor necrosis factor alpha and CD54 expression were also present. Similar phenomena were absent or substantially reduced in older infected but asymptomatic animals. These findings suggest that this murine model is suitable both for determining precise pathogenetic features of the cerebral form of the disease and for evaluating circumventive interventions.

Reversal of type 2 diabetes in mice by products of malaria parasites: I. Effect of inactivated parasites.

C57BL/KsJ-db/db and C57BL/KsJ-ob/ob mice are good models for studies on human obesity and type 2 diabetes. We have previously shown that infection with blood-stage malaria or injection of extracts from malaria-parasitized red blood cells induces hypoglycemia in normal mice and normalizes hyperglycemia in mice made moderately diabetic by streptozotocin. In the present study, we show that a single intravenous (IV) injection of Formalin-fixed Plasmodium yoelii YM (FFYM) preparation decreases blood glucose in db/db mice from an initial value of 19 mmol/L to a normal value of 7 mmol/L (P < .0001) for at least 24 hours and reduces food intake. Plasma insulin concentrations in db/db mice were not altered. FFYM was also active in normal and ob/ob mice, an effect associated with an increase in plasma insulin. Although the rate of weight gain in lean ob/+ and lean db/+ was not altered by this treatment, there was a significant reduction in weight gain in db/db and ob/ob mice (P < .001). We suggest that malaria-derived molecules, when fully characterized, may provide structural information for the development of new agents for the management of type 2 diabetes.

DNA vaccination favours memory rather than effector B cell responses.

Following priming and boosting of mice with a DNA vector pEE6DeltaS-hCGss expressing sequences encoding a transmembrane version of the beta-chain of human chorionic gonadotropin (hCGbeta), we failed to detect appreciable levels of specific antibody. However, subsequent challenge with hCG protein in Ribi adjuvant elicited a strong and rapid secondary immune response. This response was of comparable magnitude to that produced following priming, boosting and challenge with protein in adjuvant. Thus, DNA vaccination with this vector is as efficient in generating B cell memory as is conventional immunization, but the memory generation occurs in the absence of an overt effector response. Despite an overall similar level of specific antibody, the DNA-vaccinated mice produced hCG-specific antibodies biased towards IgG2a and IgG2b isotypes, whereas the protein-vaccinated mice produced higher levels of IgG1 antibodies. Both Th1 and Th2 cytokines (interferon-gamma (IFN-gamma) and IL-4) were lower in the spleens of the DNA-immunized animals compared with the protein-Ribi-immunized animals, possibly suggesting a different level of helper T cell response to the two different modes of immunization.