Usefulness of acute phase proteins for monitoring development of hepatocellular carcinoma in hepatitis B virus carriers.

BACKGROUND: Serum levels of acute phase proteins (APP) have been used to diagnose and follow up treatment of liver diseases. This study was carried out to determine the usefulness of APP to predict development of hepatocellular carcinoma (HCC) among Hepatitis B virus (HBV) carriers. STUDY DESIGN: In a prospective study, serum haptoglobin, transferrin and alpha 2 macroglobulin levels of 55 subjects consisting of 20 HBV carriers, 18 HBV-positive HCC patients and 17 controls were determined using single radial immunodiffusion. RESULT: The mean levels of haptoglobin were 141.75mg/dl+/-133.76, 97.11mg/dl+/-92.62,161.59mg/dl+/-146.86 for HBV carriers, HBV-positive HCC and controls respectively. The mean transferrin levels for HBV carriers, HBV-positive HCC and controls were 166.4mg/ dl+/-88.31, 140.0mg/dl+/-68.73 and 270.35mg/dl+/-122.79 respectively while similar values for alpha 2 macroglobulin were 195.4mg/dl+/-93.86, 189.83mg/dl+/-77.19 and 127.53mg/dl+/-43.29. No significant difference in the mean serum haptoglobin levels of HBV carriers and HBV-positive HCC (p=0.526), HBV carriers and controls (p= 0.883) and HBV-positive HCC and controls (p=0.295). The difference between the mean serum transferrin levels was insignificant between HBV carriers and HBV-positive HCC, p= 0.671, but was significant between HBV carriers, and HBV-positive HCC compared with controls, (p=0.005 and 0.000 respectively). No significant difference in alpha 2 macroglobulin between HBV carriers and HBV-positive HCC, (p=0.972), but the differences were significant between HBV carriers, and HBV-positive HCC and controls, (p=0.024 and 0.048 respectively). CONCLUSION: Haptoglobin, alpha 2 macroglobulin and transferrin lack predictive value for development of HCC in HBV carriers. Reduced transferrin and increased alpha 2 macroglobulin in HBV carriers might suggest active liver disease.

Pregnancy impairs resistance of C57BL/6 mice to Leishmania major infection.

To determine if gestational factors affect the severity of L. major infection, this study assessed the levels of IL-4 mRNA and IFN-gamma mRNA in popliteal lymph node cells of pregnant C57BL/6 mice mated at 5 hours, 16 hours and 15 days post L. major infection using PCR. Infected pregnant C57BL/6 mice developed larger cutaneous footpad lesions compared with non-pregnant infected C57BL/6 mice. The resolution of footpad lesions commenced after 8th week in C57BL/6 mice mated at 16 hrs post L. major infection but 12 weeks in C57BL/6 mice mated at 5 hrs and 15 days post L. major infection. C57BL/6 mice that were infected 20 days post partum resolved L. major infection effectively. But, the lesions in infected pregnant C57BL/6 mice and infected non-pregnant C57BL/6 mice were not as large as in susceptible BALB/c mice. The mean litter weights were similar in pregnant infected C57BL/6 mice mated at different stages of L. major infection but were slightly lower than weights of litters from pregnant uninfected C57BL/6 mice. In 5 days infected pregnant C57BL/6 mice, the levels of IFN-gamma were raised compared with the levels of IL-4 but those mated at 15 days post L. major infection had highest level of IFN-gamma mRNA. In 10 days pregnant infected C57BL/6 mice, levels of IL-4 were raised compared with IFN-gamma but mice mated at 16 hrs post L. major infection had highest level of IL-4. In 15 days pregnant infected mice, the levels of IL-4 were higher than IFN-gamma irrespective of the stage of L. major infection when the mice were mated. Mice infected with L. major 20 days post-partum produced more IFN-gamma than IL-4 from 16 hrs post L. major infection onwards. It may be concluded that increased IL-4 in pregnant infected C57BL/6 mice impairs the resistance of C57BL/6 mice to L. major infection especially in mice that were pregnant before effective immunity (5 hours post L. major infection) is mounted against L. major infection.

Interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) in pregnant C57BL/6 mice infected with L. major at different gestational periods.

BACKGROUND AND OBJECTIVE: To assess if gestational factors affect the resistance of C57BL/6 mice to L major infection, this study determined the levels of IL-4 and IFN-gamma in popliteal lymph node cells of pregnant C57BL/6 mice infected with L. major at 16 hours, 5 days-, 10 days- and 15 days- post plug by PCR, ELISA and BIOASSAY. DESIGN/SETTING: Experimental. RESULTS: Infected pregnant C57BL/6 mice developed larger cutaneous footpad lesions compared with non-pregnant C57BL/6 mice (that showed signs of resolution 7-10 weeks after infection). But, the lesions in infected pregnant C57BL/6 mice and infected non-pregnant C57BL/6 mice were not as large as in susceptible BALB/c mice. The mean litter weight was also reduced in pregnant infected C57BL/6 mice particularly in the groups infected at later stages of pregnancy (day 10- and day 15-post plug). The levels of both IL-4 and IFN-gamma increased with gestation in pregnant infected C57BL/6 mice compared with pregnant non-infected group, while only IL-4 was raised in pregnant infected mice compared with infected non pregnant mice. CONCLUSIONS: It may be concluded that increased IL-4 in pregnant infected C57BL/6 mice caused the transient susceptibility to L major infection while reduced litter weight was associated with increased IFN-gamma. These effects were pronounced in C57BI/6 mice infected with L major in late pregnancy.

Circulating immune complexes, immunoglobulin classes (IgG, IgA and IgM) and complement components (C3c, C4 and Factor B) in diabetic Nigerians.

OBJECTIVE: To evaluate serum levels of circulating immune complexes (CICs), immunoglobulin classes (IgG, IgA and IgM) and Complement Components (C3c, C4 and Factor B) in Nigerians with Type 1 or Type 2 diabetes mellitus. DESIGN: Case control study. SETTING: University College Hospital, Ibadan, Oyo State, Nigeria. SUBJECTS: Forty-two subjects with diabetic mellitus (17 Type 1 D. M. and 25 Type 2 D. M.) and 21 apparently healthy control subjects. INTERVENTION: Serum level of CICs was measured by polyethylene glycol precipitation method while single radial immunodiffusion method was used to measure serum levels of immunoglobulins and complement components. RESULTS: Only CICs were significantly higher in Type 1 diabetic subjects compared with the controls whereas CICs C3c, C4 and IgM were significantly increased in Type 2 diabetic subjects compared with the controls. The levels of CICs, C3c and IgM were significantly elevated in Type 2 diabetics compared with Type 1 diabetics. CONCLUSION: CIC concentrations may serve as a useful index of depressed host defences usually associated with diabetics mellitus and that humoral immunity is deranged more in Type 2 diabetics compared with Type 1 diabetics. Probably as a result of hyperinsulinaemia associated with insulin resistance.

Leucocyte phagocytosis and circulating immune complexes in mothers after child birth.

Leucocyte phagocytosis and level of circulating immune complexes (CICs) were measured in pregnant women at the three trimesters of pregnancy and in mothers 24-48 hours after child birth to evaluate the influence of pregnancy on leucocyte functions. The mothers were divided into gestation groups depending on the ages of their pregnancies as 1st trimester, 2nd trimester and 3rd trimester. The result shows that total white blood cell count (WBC), lymphocytes (B- and T-lymphocytes) and circulating immune complexes were high in pregnant women (independent of gestational ages) and mothers 24-48 hrs after child birth compared with non-pregnant controls. In contrast, mean percentage migration index (% M.I.), percentage candidacidal index (% C. I) and H2O2 production were reduced in pregnant women and mothers after child birth compared with non-pregnant controls. When test groups (pregnant women and mothers after child birth) were compared, women after child birth had least mean % C.I and %M.I while pregnant women at 1st trimester had highest mean %C. I., %M. I. and highest level of H2O2 production. Within the gestational groups, pregnant women at 3rd trimester had higher %M. I. compared with those in 2nd trimester while those in 2nd trimester had lower %M. I., %C. I. and H2O2 production compared with women at 1st trimester. Our finding shows that cell mediated immune responses vary between trimesters therefore susceptibility of pregnant women to different pathogens may vary with gestation.

Malaria specific-IgG, inter-pregnancy intervals, birth weights and body mass index in cases of asymptomatic malaria parasitaemia.

BACKGROUND & OBJECTIVES: Considerations of both inter-pregnancy intervals and malaria parasitaemia may help in understanding some aspects of susceptibility and pregnancy outcomes in malaria endemic areas. METHODS: Pregnant women with asymptomatic malaria parasitaemia were recruited and divided into groups based on their inter-pregnancy intervals and malaria specific-IgG, body mass index, and birth weights were studied in the groups. RESULTS: The results showed that the P. falciparum specific-IgG concentration (f=3.52, p<0.02), malaria parasites density (f=6.44, p<0.001) and birth weights (f=7.36, p<0.001) were significantly different amongst the groups with varying inter-pregnancy intervals. In addition, different levels of associations between variables such as 'inter-pregnancy intervals vs P. falciparum specific-IgG concentration' (r = 0.23, p<0.05); 'malaria parasites density vs birth weight' (r = -0.84, p < 0.01) was observed. INTERPRETATIONS & CONCLUSION: This study suggests that inter-pregnancy intervals could be one of the factors influencing dynamic serum concentrations of P. falciparum specific-IgG while malaria parasitaemia could be one of the factors affecting birth weights. Hence, observance of inter-pregnancy intervals has its own implications in malaria endemic areas.

The effects of ageing on the immune response to Schistosoma haematobium and hookworm by measuring circulating immune complexes, C3, IgG, IgA and IgM levels in residents of Omi dam area of Kogi State, Nigeria.

In this study, the effects of infestation (with Schistosoma haematobium or hookworm) during host ageing on the serum levels of circulating immune complexes (CIC), C3, IgG, IgA and IgM were examined in residents of Omi dam area of Kogi state, Nigeria. S. haematobium-infested and hookworm-infested individuals showed no significant alteration in the levels of CIC, C3, IgG, IgA and IgM in comparison with controls. These levels were the same in infested subjects and controls even when the patients were pooled. Infested old people had the same concentrations of serum CIC, C3 IgG and IgM in comparison with infested young people but IgA levels were higher in the aged group (t=2.100; P<0.05); and were significantly correlated with age (r=0.301; P<0.05). No significant increase in CIC levels with rising age (r=0.123; P>0.20) was observed in the overall population of infested subjects; and infestation in old age did not alter CIC, C3, IgG, IgA and IgM levels in comparison with uninfested young people. For the uninfested, IgG, IgA and IgM values were similar in the aged and the young but the levels of CIC were higher (t=2.156; P<0.05; r=0.280; P<0.05) and C3 lower (t=3.313; P<0.01; r=-0.236; P>0.10) in the aged. The results of this study suggest that the elevated CIC levels found in old people is age-related; and that the contribution of parasitic infestation to these raised levels is uncertain.

Complement levels and leucocyte phagocytosis in newborn babies.

Newborn babies face higher risk of infection than adults, but the immunological basis of this observation is yet to be fully explained particularly in babies of different gestations and birth weights. Sixty-two (62) adults, 55 full-term babies, 18 low birth weight babies and 44 normal birth weight babies were considered for the study. B-lymphocytes and T-lymphocytes were enumerated by EAC-rosette and E-rosette respectively. Leucocyte migration and intracellular killing were assessed by percentage migration index (%M.I), percentage Candidacidal index (%C.I) and bacterial stimulated nitroblue tetrazolium (NBT) dye reduction index (%NBT) respectively. Also, serum levels of C3 and C5 were measured by single radial immuno-diffusion method. Percentage T cell, C3, C5, %NBT and %C.I were lowest in low birth weight babies but % B cell was lowest in full term babies while normal birth babies had least %M.I. The present study suggests that gestational age and birth weight affect different aspects of immune response.

Prevalence of asymptomatic malaria parasitaemia amongst pregnant women.

Two hundred and forty-six apparently healthy pregnant women aged 19-40 years, without symptoms were recruited (147 recruited during the dry season and 99 recruited during the rainy season) for the present study. Blood examinations for malaria parasites, Plasmodium falciparum specific-IgG concentration and serological reactivity with P. falciparum-histidine rich protein-2 (HRP-2) antigens were conducted on all the pregnant women during the dry and rainy seasons of the year. During the dry season, 109 (74%) of the recruited pregnant women without symptoms had P. falciparum parasitaemia, while 79 (80%) of the recruited pregnant women without symptoms had P. falciparum parasitaemia during the rainy season. However, the P. falciparum malaria parasites density was significantly raised during the dry season compared with that of in the rainy season (p < 0.05). Serological analysis with P. falciparum histidine rich protein-2 antigen (HRP-2) showed 108 (73%) and 71 (77%) of the pregnant women without symptoms as seropositive during the dry and rainy seasons respectively. The P. falciparum specific-IgG concentration was similar during both seasons in the HRP-2 seropositive pregnant women without symptoms (p > 0.05). The results showed no seasonal tide in the incidences of asymptomatic P. falciparum parasitaemia; however, the significantly raised parasitaemia during the dry season may suggest possible increased parasites tolerance. The P. falciparum specific-IgG concentration during both seasons may not be the primary effector mechanism offering tolerance in asymptomatic parasitaemia in pregnant women.

Serum immunoglobulins, total protein and albumin levels during UniplantR use by Nigerian women.

The effects of UniplantR (a new, long-acting, 19-nor-progesterone derivative contraceptive) on serum immunoglobulins, albumin and total proteins were determined in Nigerian women during one year of use. Blood samples were collected prior to implant insertion and then at the third, sixth and twelfth months of use. All volunteers were in the reproductive age, healthy and had no contraindications to hormonal contraception. The mean levels of IgG (+/- SD) increased from pre-insertion to the twelfth month. When compared with the pre-insertion level (1,393.93 +/- 93.51 mg/dL), there are statistically significant increases in the mean values of IgG at three (1,457.19 +/- 78.41 mg/dL, p < 0.05), six (1,458.12 +/- 65.26 mgd/L, p < 0.05) and 12 months (1,499.56 +/- 87.60 mg/dL, p < 0.001). There were no statistically significant changes observed in the mean serum levels of IgA, IgM and total proteins during twelve months of implant use. These results indicate that while Uniplant does not seem to alter the levels of IgA, IgM, albumin and total proteins over a period of twelve months, it may induce significant increase in IgG levels. The raised mean serum levels of IgG may suggest an improved humoral immunity of Uniplant--a change that is potentially beneficial.

Leucocyte migration inhibitory factor assay in Nigerians with urinary schistosomiasis.

Schistosome antigens (soluble egg antigen and adult worm antigen) and non-schistosome antigens (Bacille Calmette Guerin vaccine antigen and measles virus vaccine antigen) were used to assay for leucocyte migration inhibitory factors (LMIF) in Nigerian children with and without Schistosoma haematobium infection. The severity of S. haematobium infection was graded into light infection (1-49 eggs/ 10 ml urine) or heavy infection (more than or equal to 50 eggs/10 ml urine). The mean percentage migration indices were significantly reduced in heavily infected urinary schistosomiasis (USS) subjects compared with the controls or lightly infected subjects when non-antigens were used to stimulate LMIF production. When Schistosome antigens were used to stimulate LMIF production, there were no significant decreases in the % migration indices in heavily infected USS subjects compared with the light USS subjects. The mean percentage migration index was significantly increased when schistosome antigen was used to stimulate LMIF production in treated USS subjects compared with untreated USS subjects but the increase was not significant when non-schistosome antigen was used. The conclusion that could be drawn from this study is that LMIF assay using schistosome antigen(s) has epidemiological value in schistosomiasis.

Complement haemolytic activity, circulating immune complexes and the morbidity of sickle cell anaemia.

The aim of this study was to find out if the number of crises and complications of sickle cell anaemia (SCA) relate to complement function, or the levels of circulating immune complexes (CIC), complement factor B (Bf), C3 and C4. In 73 steady-state HbSS patients and 50 HbAA control subjects, we determined the haemolytic activity of the alternative pathway of complement (AP50), of the classical pathway (CH50); and the serum concentrations of Bf, C3, C4 and CIC. By clinical examination of each patient and review of the medical records, we determined the number of complications of SCA which had occurred and the mean number of crises per year over a minimum period of 3 years. The mean+/-SD AP50 for the patients (14+/-2 U/ml) was significantly lower than the control value of 16+/-3 U/ml (p<0.001). AP50 had a significant inverse correlation with the number of crises (r=-0.30, p<0.02). Mean+/-SD CIC in patients (0.45+/-0.38 g/l) was significantly higher than in controls: 0.24+/-0.15 g/l (p<0.002). CIC showed a significant direct correlation with the number of complications of SCA (r=+/-0.28, p<0.02). Mean+/-SD Bf in SCA patients (0.19+/-0.09) was higher than in controls (0.17+/-0.05). The difference reached marginal statistical significance (p=0.049). SCA patients and controls had no significant differences in CH50, C3 and C4. These parameters and Bf did not correlate with either the number of crises or complications. The mechanisms underlying the correlations observed in this study are yet to be fully elucidated.

Levels of complement components, immunoglobulins and acute phase proteins in plasma during aging in Nigeria.

Plasma samples from Nigerians aged 6-95 years were examined for their content of complement components (C3, C4, factor B-Bf), immuloglobins (IgG, IgA, IgM IgD) and acute phase proteins (transferrin, albumin, C-reactive protein--CRP, alpha-2-macroglobulin). Albumin, was estimated colorimetrically and the other components by the single radial immunodiffusion techniques. No significant age-related changes in mean values of the four immunobulins and the four acute phase proteins could be demonstrated. Also, the mean values for C3 and Bf did not change significantly with age but C4 values rose significantly with increasing age (r -0.232: P < 0.01).

Humoral immunity in Nigerians with Schistosoma haematobium infection.

Circulating soluble immune complexes (CIC), immunoglobulin classes (IgG, IgA, and IgM), acute phase proteins (alpha-l-fetoprotein and alpha-1-acid glycoprotein) and complement factors (C4 and C3c) levels were determined in 52 Nigerian patients with urinary schistosomiasis (USS) and 39 healthy subjects by polyethylene glycol precipitation and single radial immunodiffusion methods respectively. A considerably higher proportion of USS patients than the controls had elevated levels of soluble immune complexes. IgM, IgA and C3c levels were significantly higher in USS patients than the controls. However, serum concentration of C4 was significantly reduced in USS patients than the controls while that of IgG was not significantly reduced in these patients. The mean levels of alpha-l-fetoprotein and alpha-l-acid glycoprotein were comparable in the USS subjects and the controls. Correlation analysis showed association between S. haematobium egg numbers with serum concentration of IgM, C3c and CIC. These observations could be the results of increased rate of CIC formation, and inflammatory responses to schistosome antigens. The study concluded that IgM associated-CIC may be responsible for the immunopathology of chronic schistosomiasis.

Leucocyte migration inhibition factor (L-MIF) in malnourished Nigerian children.

Leucocyte Migration Inhibition Factor (L-MIF) was measured in 41 children with marasmus, 19 with kwashiorkor, 5 with marasmic-kwashiorkor and 35 well-fed healthy children serving as controls. For L-MIF assay, two different antigens (live attenuated measles virus vaccine and diptheria pertussis tetanus (DPT) vaccine were used. Percentage migration indices obtained with the two antigens were significantly higher in the malnourished than in the well-fed healthy sex and age-matched controls (P < 0.01). The total serum protein and albumin concentrations were significantly reduced in the malnourished children compared with the controls (P < 0.01). Mean total leucocyte numbers were not significantly different in marasmic and marasmic-kwashiorkor children compared with the controls (P > 0.21).

Concurrent infections and neutrophil phagocytic function in Nigerians with urinary schistosomiasis.

Bacteria infections, parasitic infections and neutrophil phagocytic function were assessed in 60 urinary schistosomiasis (USS) subjects and 36 healthy controls. Only Salmonella was isolated from the blood samples of few USS subjects (3%) and controls (1%). The species of bacteria detected in the urine of USS and control subjects are Escherichia coli, Salmonella, Staphylococcus aureus, Streptococcus faecalis and Klebsiella. There was no significant difference in the proportion of the USS subjects with bacteriuria compared with the controls (X2 = 0.20, P > 0.20). Higher proportions of USS subjects compared with the controls were concurrently infected with Ascaris lumbricoides, hookworms, Giardia lambila and Taenia solium. In contrast, significantly higher proportion of the control subjects were concurrently infected with Plasmodium falciparum and Schistosoma mansoni. There was no significant reduction in neutrophil phagocytic function of USS subjects compared with the controls (P > 0.49). This study suggests that S. haematobium protects its host from certain blood dwelling parasites and that Nigeria USS subjects expressed adequate neutrophil phagocytic function. These may explain the absence of clinical manifestations of bacterial and viral infections in these subjects.