Urinary Antibody Research Articles

Diagnosis of hepatitis C infection by Elisa detection of urinary antibody

Diagnosis of hepatitis C infection by Elisa detection of urinary antibody

Anti-DNA antibodies in the urine of lupus nephritis patients

It has previously been reported that patients with systemic lupus erythematosus (SLE) and glomerulonephritis do not have anti- (deoxyribonucleic acid) DNA antibodies in their urine. This finding was attributed to specific entrapment of anti-DNA antibodies by the immune complexes in the glomerular capillary walls. This phenomenon has been re-investigated as part of a study of the use of desoxyribonuclease 1 (DNase 1) to treat lupus nephritis (LN). For this purpose an ELISA was developed for the detection of anti-DNA antibodies in urine. It was found that such an assay was very susceptible to the presence of DNase in urine which destroys the antigen coating the plates and gives rise to false negative results. For this reason, it is essential that all tests for anti-DNA antibodies in the urine are carried out in the presence of EDTA to inhibit the endogenous DNase 1 activity. Using this assay to test the urine from 24 patients with LN and non-selective proteinurea, it was found that they all contained anti-DNA antibodies. The amount of anti-DNA antibodies detected in the urine was compared with that expected by calculations from the anti-DNA antibody titre in the serum and total immunoglobulin levels in serum and in urine. It showed that in 20 patients there was neither specific entrapment nor specific excretion of anti-DNA in urine, only the expected amount of leakage. In only three patients was any appreciable entrapment demonstrated and in only one, any excess excretion. It is suggested that the failure to detect anti-DNA antibodies in the urine in the previous work was due to failure to inhibit the endogenous urinary DNase. It remains to be determined whether the retention of anti-DNA antibodies or excessive secretion is correlated with clinical phases of LN.

Urinary Antibody Level and Survival in Bacteriuric Institutionalized Older Subjects

OBJECTIVE: In a previous study, elevated urine antibody levels in institutionalized subjects with asymptomatic bacteriuria were associated with decreased survival. This study was undertaken to confirm the previous observation in a prospective study in a larger cohort and to explore selected other variables that may be associated with survival. DESIGN: A prospective, 24-month, observational cohort study. SETTING: Three large nursing homes in Winnipeg, Manitoba. PARTICIPANTS: Permanent residents were identified by initial screening urine cultures and subjects with bacteriuria were enrolled. The median age of subjects was 76 years, 51% were women, and the median duration of residence before enrollment was 26 months. Subjects were highly functionally impaired. MEASUREMENTS: Monthly urine specimens were collected for culture, leukocyte count, and urine antibody. Serum specimens for antibody to uropathogens and IL6 were obtained at enrollment and every 6 months. Anthroporphometric tests of nutritional status and functional and mental status were also measured every 6 months. Residents were stratified as having elevated or not elevated urine antibody, based on the initial urine specimen. The mean urine antibody for all of each subject's specimens was also calculated, and subjects were stratified as low, intermediate, or elevated urine antibody. Outcomes measured included mortality, infection and antibiotic use, and functional, mental, and nutritional decline. RESULTS: Ninety-eight bacteriuric subjects were enrolled in the study; 34 (35%) had elevated urine antibody and 64 (65%) had not elevated urine antibody. The two groups did not differ in demographic features, co-morbidities, functional status, medication use, or infecting organisms. Survival was significantly (P <.001) poorer in the group with elevated urine antibody. At 24 months, 35% of subjects with an elevated urine antibody were alive compared with 75% with a low urine antibody level. This survival difference was consistent when the two groups were stratified by sex, institution, and presence of a chronic indwelling catheter. Subjects with elevated urine antibody had no evidence for accelerated functional or nutritional decline during the study period compared with the group with low urine antibody. These subjects did, however, have an increased incidence of episodes of symptomatic urinary infection and infections at non-urinary sites. CONCLUSIONS: Older, bacteriuric, institutionalized subjects with elevated urine antibody had decreased survival rates compared with subjects with lower urine antibody levels. There is no clinical evidence to support accelerated decline caused by chronic infection to explain this observation. Urine antibody may be a marker for immune dysregulation, which precedes death in older impaired subjects. J Am Geriatr Soc 46:947–953, 1998.

Urinary Antibody Level and Survival in Bacteriuric Institutionalized Older Subjects

Urinary Antibody Level and Survival in Bacteriuric Institutionalized Older Subjects

Assessment of mucosal immune response in genitourinary tract using urine.

A novel method to assess mucosal immune response in the genitourinary mucosa after immunization with a mucosal vaccine has been developed. In this method, secretory IgA antibody is measured by a highly sensitive enzyme immunoassay (immune-complex transfer enzyme immunoassay) using urine as a specimen. The urinary IgA antibody response could be detected by the immune-complex transfer enzyme immunoassay. In contrast, a conventional enzyme immunoassay (enzyme-linked immunosorbent assay (ELISA)) could not detect this response because of its low sensitivity. Because urine samples can be collected easily and nontraumatically, not only from experimental animals but also from humans, both males and females, the present method may be applicable for assessing the protective efficacy of candidates for mucosal vaccines against sexually transmitted microorganisms, such as human immunodeficiency virus. Furthermore, the usefulness of this method for novel mucosal vaccine formulae was shown for a model in which vaccine antigen and Bordetella pertussis adjuvant were adsorbed onto CaCO, and enclosed in enteric coated capsules.

Diagnostic modalities for the detection of Helicobacter pylori.

Numerous techniques are available to detect Helicobacter pylori. The first were invasive methods based on biopsy specimens. While these methods still constitute the reference, noninvasive tests are in continuous development. After laboratory-based serology and (13)C-urea breath tests, an antigen stool test is now available, as well as tests to detect antibodies in saliva, urine and whole blood tests which can be used in the physician's office. Most of the tests are adapted to a particular situation. However, more evaluation is needed, especially in pediatric and geriatric populations.

Urinary loss of immunoglobulin G anti-F(ab′) 2 and anti-DNA antibody in systemic lupus erythematosus nephritis

The objective of this study was to determine whether the low levels of serum immunoglobulin G (IgG) anti-F(ab′) 2 seen in some patients with active systemic lupus erythematosus (SLE) were directly related to the deposition of antibody with this specificity in the kidney or alternatively to the urinary loss of IgG anti-F(ab′) 2. Serum Levels of IgG anti-F(ab′) 2, anti-tetanus toxoid, and anti-ds DNA antibody were measured in parallel with urinary excretion of these same 3 antibodies in 28 patients with SLE nephritis and in 28 control patients with other forms of chronic kidney disease. Low levels of both serum IgG anti-F(ab′) 2 or anti-tetanus antibody appeared to correlate with increased levels of urinary loss of these same antibodies in some patients with SLE and in control subjects with kidney disease. However, urinary loss could not account for low serum levels of either IgG antibody in many subjects. Quantitative 24-hour urinary losses of IgG anti-F(ab′) 2 and anti-DNA were much higher in patients with SLE than in control subjects with kidney disease ( P < .05), whereas amounts of IgG urinary loss of anti-tetanus were similar in patients with SLE and in control subjects. In nearly 1 third of SLE nephritis patients, 13% to 53% of total excreted urinary IgG showed anti-DNA enzyme-linked-immunosorbent assay reactivity. Urinary IgG in many patients with SLE showed both anti-DNA and anti-F(ab′) 2 reactivity, but dual anti-DNA/F(ab′) 2 specificity was more pronounced in affinity-isolated serum IgG anti-DNA or anti-F(ab′) 2 than in excreted urinary IgG molecules. The affinity of urinary IgG for either DNA or F(ab′) 2 was much lower than the same antibody activities measured either in serum or in kidney biopsy eluates. When the relative affinity of anti-DNA antibody in serum, urine, and kidney biopsy eluate was measured in parallel, the highest affinity antibody was found in kidney biopsy eluates, followed by serum antibody with urine antibody affinity showing the lowest values. These findings suggest a relative concentration of the highest affinity, doubly reactive IgG anti-DNA/F(ab′) 2 in SLE kidney tissues during SLE nephritis and implicate this process as an important factor in ongoing tissue damage.

Urinary antibody level and survival in bacteriuric institutionalized older subjects.

In a previous study, elevated urine antibody levels in institutionalized subjects with asymptomatic bacteriuria were associated with decreased survival. This study was undertaken to confirm the previous observation in a prospective study in a larger cohort and to explore selected other variables that may be associated with survival. A prospective, 24-month, observational cohort study. Three large nursing homes in Winnipeg, Manitoba. Permanent residents were identified by initial screening urine cultures and subjects with bacteriuria were enrolled. The median age of subjects was 76 years, 51% were women, and the median duration of residence before enrollment was 26 months. Subjects were highly functionally impaired. Monthly urine specimens were collected for culture, leukocyte count, and urine antibody. Serum specimens for antibody to uropathogens and IL6 were obtained at enrollment and every 6 months. Anthroporphometric tests of nutritional status and functional and mental status were also measured every 6 months. Residents were stratified as having elevated or not elevated urine antibody, based on the initial urine specimen. The mean urine antibody for all of each subject's specimens was also calculated, and subjects were stratified as low, intermediate, or elevated urine antibody. Outcomes measured included mortality, infection and antibiotic use, and functional, mental, and nutritional decline. Ninety-eight bacteriuric subjects were enrolled in the study; 34 (35%) had elevated urine antibody and 64 (65%) had not elevated urine antibody. The two groups did not differ in demographic features, co-morbidities, functional status, medication use, or infecting organisms. Survival was significantly (P < .001) poorer in the group with elevated urine antibody. At 24 months, 35% of subjects with an elevated urine antibody were alive compared with 75% with a low urine antibody level. This survival difference was consistent when the two groups were stratified by sex, institution, and presence of a chronic indwelling catheter. Subjects with elevated urine antibody had no evidence for accelerated functional or nutritional decline during the study period compared with the group with low urine antibody. These subjects did, however, have an increased incidence of episodes of symptomatic urinary infection and infections at non-urinary sites. Older, bacteriuric, institutionalized subjects with elevated urine antibody had decreased survival rates compared with subjects with lower urine antibody levels. There is no clinical evidence to support accelerated decline caused by chronic infection to explain this observation. Urine antibody may be a marker for immune dysregulation, which precedes death in older impaired subjects.

HCV antibodies in saliva and urine

Infection with hepatitis C virus (HCV) is usually established by detection of serum antibodies (anti-HCV). This study was conducted in order to evaluate whether saliva and urine may substitute serum for anti-HCV detection. Serum, saliva, and urine were obtained simultaneously from 141 patients with a variety of liver diseases and from 52 patients with autoimmune diseases (systemic lupus erythematosus n = 27 and rheumatoid arthritis n = 25). The cell free fraction of saliva and urine samples was tested for anti-HCV using a modification of a serum anti-HCV kit. Western blot analysis was used as a confirmation method. Of the patients with liver diseases, 73 were anti-HCV-seropositive. Salivary and urinary anti-HCV could be detected in 66 (90%) and 36 (49%) of the anti-HCV-serpositive patients, respectively. The presence of anti-HCV in saliva or urine was not related to the severity of liver disease. All the anti-HCV-seronegative liver patients were negative for salivary anti-HCV and 22 (32%) had urinary anti-HCV. The patients with autoimmune diseases were all anti-HCV-seronegative. None had detectable salivary anti-HCV while 33 (63%) were positive for urinary anti-HCV. Western Blot analysis confirmed the presence of anti-HCV in all serum and saliva samples tested but only in 2/12 urine samples. The results suggest that saliva, but not urine, may serve as a substitute for serum for the determination of anti-HCV positivity. J. Med. Virol. 55:24–27, 1998. © 1998 Wiley-Liss, Inc.

HCV antibodies in saliva and urine

Infection with hepatitis C virus (HCV) is usually established by detection of serum antibodies (anti-HCV). This study was conducted in order to evaluate whether saliva and urine may substitute serum for anti-HCV detection. Serum, saliva, and urine were obtained simultaneously from 141 patients with a variety of liver diseases and from 52 patients with autoimmune diseases (systemic lupus erythematosus n = 27 and rheumatoid arthritis n = 25). The cell free fraction of saliva and urine samples was tested for anti-HCV using a modification of a serum anti-HCV kit. Western blot analysis was used as a confirmation method. Of the patients with liver diseases, 73 were anti-HCV-seropositive. Salivary and urinary anti-HCV could be detected in 66 (90%) and 36 (49%) of the anti-HCV-seropositive patients, respectively. The presence of anti-HCV in saliva or urine was not related to the severity of liver disease. All the anti-HCV-seronegative liver patients were negative for salivary anti-HCV and 22 (32%) had urinary anti-HCV. The patients with autoimmune diseases were all anti-HCV-seronegative. None had detectable salivary anti-HCV while 33 (63%) were positive for urinary anti-HCV. Western Blot analysis confirmed the presence of anti-HCV in all serum and saliva samples tested but only in 2/12 urine samples. The results suggest that saliva, but not urine, may serve as a substitute for serum for the determination of anti-HCV positivity.

HTLV-I-associated myelopathy associated with multi-organ inflammatory disease: a case report

We report a 73-year-old man with human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (HAM) complicated with multi-organ inflammatory disease, including Sjögren's syndrome, interstitial cystitis, and uveitis. The presence of HTLV-I proviral DNA in peripheral blood mononuclear cells (PBMC), cerebrospinal fluid, salivary gland, mucosa of urinary bladder, and aqueous humor was confirmed by polymerase chain reaction using HTLV-I pX region primer. Western blot analysis revealed the presence of anti-HTLV-I antibodies in serum, CSF, saliva, and urine, suggesting replication of HTLV-I in each tissue. A high load of HTLV-I proviral DNA (20 copies out of 100 PBMC) was present, associated with increased spontaneous proliferation of peripheral blood lymphocytes (24 747 cpm). Our results suggest that the high load of HTLV-I in patients with HAM may potentially induce systemic inflammation in several organs.

Urine-based diagnostic technologies

The worldwide dissemination of infectious agents has created a demand for simple diagnostic tests. Urine-based testing makes use of non-invasive collection of specimens, and there is no need for expensive facilities and equipment, or for highly trained personnel. As urine antibodies retain activity under normal conditions of transport and storage, such tests appear to have widespread application. Urine-based antibody tests have also indicated a compartmentalized antibody response to HIV-1 infection. Urine studies suggest that antibodies to the products of endogenous viral genes may be involved in the pathogenesis of chronic diseases of suspected viral etiology.

Diagnosis of respiratory virus infections using GACELISA of urinary antibodies

Sensitive and specific methods are needed to diagnose respiratory virus infections using body fluids such as urine that, unlike blood samples, are readily obtained by non-invasive means. Immunoglobulin G antibody capture enzyme-linked immunosorbent assays were developed for detection of antibody rises to respiratory syncytial virus and influenza A/Taiwan 3H1N1) after initial quantification and adjustment of urinary IgG concentration. Of 24 elderly subjects whose sera were assayed by the complement fixation test for antibody to RSV, seven had convalescent titres ≥ 32, and five had ≥ 4-fold rises in titre. Acute and convalescent urines for six of these seven subjects were tested for virus-specific urinary IgG by GACELISA. Four of four persons with ≥ 4-fold rises in CFT had urine ELISA convalescent to acute ratios of ≥ 1.8 whereas two subjects with convalescent CF titres > 16, but no increase in serum antibody titre, had urine convalescent/acute ratios of 1.0. Ten subjects with ≥ 4-fold rises in CFT or HI antibodies to influenza A/Taiwan had urine ELISA ratios of ≥ 1.4 when samples taken on the day of influenza vaccination and 16 days later were compared. These preliminary observations demonstrate clinically significant rises in respiratory pathogen antibody levels between acute and convalescent urine samples, provided that total urinary IgG concentrations are quantified and then standardised.

Detection of antibodies to Shigella lipopolysaccharide in urine after natural Shigella infection or vaccination.

The purpose of the present study was to explore the possibility of detecting antibodies to Shigella sonnei lipopolysaccharide (LPS) in urine after infection or vaccination. Urinary immunoglobulin A (IgA) and IgG antibodies and specific IgA secretory protein against S. sonnei LPS were measured by enzyme-linked immunosorbent assay (ELISA), after adjustment for urine concentration. A significant antibody level was defined as one above a cutoff value calculated from the geometric mean + 2 standard deviations of urinary anti-S. sonnei LPS levels in 43 healthy hepatitis B vaccinees (controls). Of 11 culture-proven cases of S. sonnei shigellosis, at convalescence 9 (82%) had significantly elevated levels of urinary antibodies to the homologous LPS. The S. sonnei conjugate vaccine, composed of S. sonnei O-specific polysaccharide covalently bound to recombinant exoprotein A of Pseudomonas aeruginosa, elicited a significant urine IgA or IgG anti-LPS response in 60% (6 of 10), 56% (9 of 16) 43% (16 of 37), and 14% (3 of 21) of the volunteers at 2 weeks, 6 weeks, 6 months, and 12 months after vaccination, respectively. The specificity of the urine antibody response to S. sonnei LPS was documented by the total lack of response in subjects who received parenteral Shigella flexneri 2a-recombinant exoprotein A conjugate (69 urine samples) or meningoccal tetravalent control vaccines (4 urine samples). All the volunteers who lacked a significant response to S. sonnei LPS in serum also lacked such response in urine samples. Seventy-four percent of the volunteers with a significant IgA or IgG anti-LPS response in serum at convalescence or 14 days after vaccination showed a similar response in urine. The ratio of the titer of secretory protein bound to IgA anti-S. sonnei LPS in urine to that in serum was 303 times higher than the ratio of anti-S. sonnei LPS total IgA titer in urine to that in serum, indicating that the urine IgA is of secretory origin. These findings suggest the possible use of urinary Shigella LPS antibodies as markers of systemic and secretory immune responses after natural infection or vaccination. At this stage, because of its limited sensitivity, the detection by ELISA of Shigella LPS antibodies in urine cannot replace the same assay in serum as a definitive test in an individual with a negative result.

5 Diagnosis of Helicobacter pylori

Several techniques have been proposed to diagnose H. pylori infection based either on the direct or indirect detection of the bacteria. The current direct methods are performed on biopsy specimens obtained at endoscopy. Histological examination must always be performed because in addition to H. pylori detection it allows observation of the lesions present. Culture, while more demanding in terms of transport conditions, is a sensitive technique which is worthwhile when the patient might be treated with antibiotics to which the bacteria may be resistant. PCR has been more recently introduced and exhibits a good sensitivity and specificity. Rapid tests, such as the rapid urease test and examination of a Gram stained biopsy smear, are convenient and cheap but of lower sensitivity. Indirect methods are based either on the serological IgG response or the labelled urea breath test. Both are sensitive. The breath test is especially adapted to post-treatment control while serology is of interest for screening patients and epidemiological studies. New techniques have recently been proposed but are not yet applicable to routine diagnosis: detection of H. pylori in faeces by culture or PCR; detection of IgG antibodies in saliva or urine.

Resolution time of Escherichia coli cystitis is correlated with levels of preinfection antibody to the infecting Escherichia coli strain

Objectives. A number of studies in animals have demonstrated that immunization against uropathogenic bacteria can lessen the severity or duration of induced urinary tract infections (UTI). This study examined whether preinfection levels of serum or urinary anti- Escherichia coli antibodies were correlated with length of time needed to resolve an induced E coli cystitis. Methods. Serum and urine anti- E coli antibody levels in 36 control and 42 vaginally immunized cynomolgus monkeys were measured by enzyme-linked immunosorbent assay. Regression analyses were used to determine correlations between resolution time and preinfection antibody level, and to estimate antibody levels that might be associated with effective resolution of an E coli UTI. Results. Linear regression analysis showed significant correlations between short resolution time and high levels of serum immunoglobulin M (IgM), urinary secretory IgA, and urinary IgG specific for the infecting E coli strain. Serum IgM and urinary IgG anti- E coli levels in monkeys that cleared infections early were significantly higher than in animals with protracted infections. Logistic regression estimated that serum IgM and urinary IgG anti- E coli levels associated with a 50% probability of accelerated clearance to be 3.3 μg/mL and 2.7 μg/24 hours, respectively. Conclusions. For this primate model, the observed correlations between short resolution time and pre-existing serum and urinary antibody suggest that antibody-mediated immunity is an important component of host defense against UTI.

Phase I Clinical Trial of Vaginal Mucosal Immunization for Recurrent Urinary Tract Infection

In a phase I clinical trial to test safety and patient acceptance 25 women with a history of recurrent urinary tract infections but no identifiable anatomic abnormality received a multivalent vaccine instilled into the vagina. The vaccine contained 6 heat-killed Escherichia coli strains and 4 nonE. coli uropathogens. Only minimal adverse reactions were observed in the 5-month period following immunization. Total vaginal and urinary IgG and IgA increased significantly (p <0.01 by repeated measures analysis of variance). Serum antibodies to some of the nonE. coli strains but not to the E. coli strains increased after vaginal immunization. While efficacy is yet to be shown, this study indicates that this vaginally applied urinary tract infection vaccine is well tolerated, and capable of increasing vaginal and urinary antibody.

Risk of preterm delivery in pregnant women with group B streptococcal urinary infections or urinary antibodies to group B streptococcal and E. coli antigens

To establish whether there is an association between preterm delivery and either group B streptococcal urinary infection or the presence of urinary antibodies to group B streptococcal or E. coli antigens. A prospective study with urine culture and antibody measurement performed at the first antenatal visit and at 28 weeks gestation. Ninewells Hospital, Dundee. Two thousand and forty-three women registering consecutively at an antenatal clinic. Delivery at less than 37 weeks gestation. No increase in preterm delivery was observed in women with positive urine cultures for group B streptococci either at booking or at 28 weeks, even when confirmed by positive repeat cultures. Preterm delivery was more common in women with elevated urinary antibodies to E. coli antigens at booking (relative risk 1.81, 95% CI 1.22-2.68, P = 0.005) and at 28 weeks (relative risk 2.36, 95% CI 1.60-3.48, P < 0.0001) and to group B streptococcal antigens at 28 weeks (relative risk 2.24, 95% CI 1.46-3.43, P = 0.0003). These data do not support previous reports that positive urine cultures for group B streptococci are associated with an increased risk of preterm delivery. Our report of an association between elevated levels of urinary antibodies and preterm delivery is a new finding consistent with the possibility that a local inflammatory response to uro-genital infection may be important in stimulating the onset of preterm labour. The results suggest that screening for urinary antibodies at 28 weeks gestation might help to identify a group of women at increased risk of prematurity.

Suppressed antibody and interleukin-6 responses to acute pyelonephritis in pregnancy

This study examined the effect of pregnancy on the host response to acute pyelonephritis. Urine and serum samples were obtained at the time of diagnosis and after two weeks, from non-pregnant and pregnant women with acute pyelonephritis. The samples were analyzed for interleukin-6 (IL-6) and specific antibody activity to antigens extracted from the Escherichia coli strain infecting each patient. The host response to infection was further quantitated as fever, C-reactive protein, and renal concentrating capacity. Acute pyelonephritis in non-pregnant and pregnant women was accompanied by a significant serum and urine antibody response. The serum antibody response was significantly lower in the pregnant group. The IL-6 levels in serum and urine at diagnosis were significantly higher in the non-pregnant compared to the pregnant women. These results demonstrate that the immunosuppression of pregnancy includes the mucosal IL-6 and specific antibody responses to acute pyelonephritis caused by E. coli.

Novel and Sensitive Enzyme Immunoassay for Urinary Antibody IgG to β-D-Galactosidase from Escherichia coli

Abstract A novel and sensitive enzyme immunoassay for antibody IgG in urine, in which the concentration of IgG is extremely low, is described. Antibody IgG to β-D-galactosidase from Escherichia coli was used as a model antibody. Anti-β-D-galactosidase IgG in urine was reacted with 2,4-dinitrophenyl-β-D-galactosidase. β-D-Galactosidase was used not only as antigen but also as label enzyme. The immune complex formed was trapped onto two colored polystyrene balls coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG. After washing, the immune complex was eluted from the colored polystyrene balls with excess of eN-2,4-dinitrophenyl-L-lysine, reacted with biocytin-S-S-(antihuman IgG γgM-chain) IgG and trapped onto two white polystyrene balls coated with streptavidin. The white polystyrene balls were washed, and the immune complex was eluted by reduction with excess of 2-mercaptoethylamine and transferred to two colored polystyrene balls coated with affinity-purified (anti-human IgG γgM-chain) IgG. β...