Cervical screening in general practice: call and recall

Regular universal screening for cervical cancer is associated with a considerable reduction in the disease. However, opportunistic screening has tended to reach groups at low risk and miss those at high risk from the disease. This study assessed the cost-effectiveness of a call and recall system for cervical screening which was set up in one general practice.

The practice age-sex register and records were used to monitor the screening status of women patients. Of the eligible population aged 36-60 years 70% were found to have been screened in the previous five years. The remainder were offered an appointment for a cervical smear and 57% attended following this invitation. Three smears out of 110 undertaken were reported as showing marked dyskaryosis or cervical intraepithelial neoplasia grade III. The estimated cost per case identified was £366.

A call system in general practice can increase the uptake of cervical screening among women at risk. It is a relatively cost-effective method of preventing cervical cancer.

     

Studies of the third component of complement in synovial fluid from arthritic patients. II. Conversion and its relation to total complement

Plasma and synovial fluid from arthritic patients were studied with antigen–antibody crossed electrophoresis for the conversion of C3. When present, C3 conversion was estimated planimetrically. The material included patients with rheumatoid arthritis and systemic lupus erythematosus as well as patients with non-rheumatoid arthritis.

C3 conversion was not found in plasma from any of the patients studied.

In non-rheumatoid synovial fluids there was no conversion in five and less than 10% in four of the samples. In rheumatoid synovial fluids C3 conversion proved significantly (P<0·01) more pronounced, the degree of conversion exceeding 10% in fourteen out of twenty-three cases. An inverse relationship was found in synovial fluid between the degree of C3 conversion on the one hand, and the total complement activity or the C3 concentration on the other.

     

A pilot study on Hla-G locus control region haplotypes and cervical intraepithelial neoplasias

Human papillomavirus (HPV) can induce cervical intraepithelial neoplasias (CIN) grades 1, 2 and 3. Untreated, these lesions may progress to cervical cancer (CC) which is the third most common cancer in women worldwide. HLA-G plays an immunotolerant role in the immune response. The aim of this study was to characterize the configuration of SNPs located at the distal promoter of HLA-G in patients with CIN2 and CIN3 and control women. The study sample was composed of 207 women as follows: 73 diagnosed with CIN2 lesions, 56 with CIN3 and 78 healthy control women. Genotyping was performed by sequence base typing. Eleven haplotype configurations subdivided in two main haplogroups (H1dist and H2dist), were characterized and compared between patients and controls. The haplotypes H1.1Dist (GAGAACGC) and H2.1Dist (AGGTACAC) were more frequent in Euro-Descendants as well as in Brazilian Mixed. Nevertheless, the haplotype H2.1Dist standed out as a susceptibility haplotype in Brazilian Mixed patients while the H1.1Dist presented a protector effect in this same ethnic group. Whether such LCR haplotype configurations can impact on HLA-G gene expression levels in women who developed cervical intraepithelial neoplasia is still unknown and it is of utmost importance that more investigation on this field be pursued.     

Autoantibodies to haemoglobin in cattle

Two cases are described of autoantibodies to haemoglobin being produced in cattle infected with Corynebacterium pyogenes. The antibody in one of these animals has been characterized and found to be a transient IgM precipitating antibody.

The reaction of the antibody with haemoglobins from other species has been tested and it has been found to react with all normal mammalian haemoglobins tested and bird haemoglobin but not fish haemoglobin.

The possible role of C. pyogenes both as a producer of haemolytic toxin and as an adjuvant in antibody production is discussed.

     

Faisabilité du frottis cervico-utérin chez les femmes séropositives pour le VIH vivant au Tchad

Cervical cancer is the leading cause of cancerrelated death in Sub-Saharan African women. HIV-infected women are at increased risk for cervical intraepithelial lesions and invasive cervical cancer. WHO guidelines for screening and treatment of precancerous cervical lesions are regularly actualized. There are no data on cervical squamous intraepithelial lesions in Chad. Between August 2013 and May 2015, screening for cervical squamous intraepithelial lesions was proposed to HIV-infected women living in Moundou (Chad). Cytology examination was performed after with Papanicolaou coloration. Three hundred and eleven HIV-seropositive women accepted the screening without refusal. Mean age of the patients was 38 years (95% Confidence Interval: 37.7–39.9). The women declared a mean of 4.1 pregnancies (range: 0–12). The patients had been followed-up for their seropositivity for 8 years (range: 0–25). All were on highly active antiretroviral therapy (HAART). Of the patients whose results were known (N = 231), 98% had a CD4 lymphocyte nadir count less than 350/mm³. Cytological results were as follows: normal smear (N = 59; 19%), inflammatory or hemorrhagic smear (N = 139; 44%), low grade squamous intraepithelial lesion (N = 58; 19%), high grade squamous intraepithelial lesion (N = 28; 9%), epidermoid carcinoma (N = 13; 4%), and uninterpretable smear (N = 14; 5%). The inflammatory lesions were due to cervicitis (N = 54), vaginosis (N = 22), and trichomonas infection (N = 3). The patients’ age, CD4 lymphocyte nadir count, and CD4 count at the time of the cervical smear were not different according to the cytological results. Only five patients had a cone biopsy. Three patients deceased during the study of whom two from a gynaecological cancer diagnosed too late. The screening of dysplasia and cervical cancer in HIV-seropositive women is possible in Chad. In our study, 13% of the women had highgrade dysplasia or carcinoma needing curative care. We also showed that simple cytology did not permit the interpretation of half of the smears. The performance of cervical smear would have increased if it had been preceded by the visualization of the cervix with coloration.     

Porcine colostral IgA and IgM antibodies to Escherichia coli and their intestinal absorption by the neonatal piglet

Antibodies against Escherichia coli 0141 and Escherichia coli 08 have been studied in porcine colostrum and serum using gel filtration and DEAE-cellulose chromatography. Immunoglobulins were assayed by radial immunodiffusion.

Immune inhibition studies with a rabbit anti-IgA-globulin serum showed that a high proportion of antibodies in colostrum measured by the antiglobulin haemagglutination test were associated with IgA. The IgA antibody could not be detected in sow serum where the antibody was almost entirely confined to IgM. Thus it appears that colostral IgA antibody was synthesized in the mammary gland.

Studies of the absorption of immunoglobulins and antibodies from colostrum in six litters of piglets showed that although IgA was absorbed from the colostrum, colostral IgA antibody to E. coli was not acquired as part of the passive immunity of the neonatal piglet.

The absorbed antibody was associated with IgM; another high molecular weight immunoglobulin 18S IgG was also absorbed.

It is suggested that structures on secretory IgA which correspond to cellular receptors for intestinal transmission may be blocked by the integration of secretory `piece'.

     

The immune response to influenza virus: II. Effect of the route and schedule of vaccination on the quantity and avidity of antibodies

Anti-haemagglutinin titres and measurements of the changes in the number of antibody molecules per millilitre (A), and in their average avidities (K) were carried out by equilibrium filtration on antisera prepared in rabbits to SW, MEL and LEE influenza viruses. These studies showed:

(1) Immunization of rabbits with influenza virus by the intravenous route induced sufficient levels of antibodies by the 5th day after vaccination for testing by equilibrium filtration, immunization by the intraperitoneal or subcutaneous routes did not induce sufficient antibody levels for testing by the 5th day.

(2) The avidity of the antibodies present at 5 days after vaccination was higher than at 10 days after vaccination.

(3) The avidity of antibodies increased during the initial response period (from the low point at 10 days) and was independent of the routes or schedules of vaccination tested.

(4) Secondary vaccination did not cause a pronounced increase in the avidity of antibodies, although a few groups of rabbits showed some further increase; tertiary vaccination caused no further increase.

(5) The number of antibody molecules produced in rabbits after vaccination with influenza virus depended on the route and schedule of inoculation; the routes could be ordered intravenous, intraperitoneal and subcutaneous in decreasing order of efficiency; multiple doses increased the number of antibody molecules by all routes.

(6) Increases in the antihaemagglutinin levels after secondary stimulation were due mainly to increases in the number of antibody molecules.

     

Effect of tissue inhibitor of metalloproteinases-3 genetics polymorphism on clinicopathological characteristics of uterine cervical cancer patients in Taiwan

Single nucleotide polymorphisms (SNPs) of tissue inhibitor of metalloproteinases-3 (TIMP-3) have been revealed to be related to various cancers. To date, no study explores the relationships between TIMP-3 polymorphisms and uterine cervical cancer. The purposes of this research were to investigate the associations among genetic variants of TIMP-3 and development and clinicopathological factors of uterine cervical cancer, and patient 5 years survival in Taiwanese women. The study included 123 patients with invasive cancer and 97 with precancerous lesions of uterine cervix, and 300 control women. TIMP-3 polymorphisms rs9619311, rs9862 and rs11547635 were checked and their genotypic distributions were determined by real-time polymerase chain reaction. It showed that women with genotypes CT/TT in rs9862 were found to display a higher risk of developing cervical cancer with moderate and poor cell differentiation. Moreover, it revealed that cervical cancer patients carrying genotypes CC in rs9619311 exhibited a poorer 5 years survival, as compared to those with TT/TC in Taiwanese women, using univariate analysis. In addition, pelvic lymph node metastasis was determined to independently predict 5 years survival in cervical cancer patients using multivariate analysis. Conclusively, TIMP-3 SNPs polymorphisms rs9619311 are related to cervical patient survival in Taiwanese women.     

Studies on the transfer of antibody-producing capacity. I. The transfer of antibody-producing cells to young animals

Normal adult spleen cells were antigenically stimulated by incubation with isolated flagella from Salmonella bacteria, and after neutralization of free antigen and washing, were transferred to normal and sublethally X-irradiated mice and rats of various ages by intraperitoneal injection. Neonatal rats and Hall Institute mice could not produce antibody following such injections, but more mature rats and Hall Institute mice, and neonatal C₃H mice, could produce antibody.

Cells from immunized spleens were secondarily stimulated in vitro and transferred to neonatal and more mature recipients. Regardless of the age of the recipients, antibody production typical of a secondary response ensued.

Cells from neonatal rat spleens were stimulated in vitro and transferred to more mature recipient rats. Some antibody production resulted.

The theoretical implications of these findings are discussed.

     

Agglutinating and non-agglutinating antibodies in rabbits inoculated with a particulate antigen (Salmonella typhimurium)

Agglutinating and non-agglutinating anti-Salmonella typhimurium antibodies were specifically purified from the sera of immunized rabbits. Both types of antibody had the same electrophoretic mobility and were localized in the IgG fraction. It was not possible to find antigenic differences between agglutinating and non-agglutinating antibodies by immunodiffusion.

Agglutinating antibody activated the complement system, while non-agglutinating antibody lacked this capacity. Only the former increased clearance of antigen from the blood. When serum samples with different antibody titres determined by agglutination (agglutinating antibody) and Coombs test (non-agglutinating antibody) were injected in mice, clearance of antigen from the blood showed changes. These results were similar to those previously observed by us when different precipitating: co-precipitating antibody ratios were used, and indicated that competition of both antibodies for the antigen depends on their respective amounts.

When mice protection tests were set up by injection of agglutinating and non-agglutinating antibody before the inoculation of 10 LD₅₀ S. typhimurium, non-agglutinating antibody was found to be less effective than agglutinating antibody.

Non-agglutinating antibody was detectable during the whole course of immunization. Its serum concentration was higher than that of the agglutinating antibody.

Non-agglutinating antibody behaves in a similar way to co-precipitating antibody. The initially proposed hypothesis that such antibodies could interfere with immunity to certain chronic infections was extended to include the non-agglutinating antibodies demonstrated here.

     

Electron microscopic studies of the antigen-antibody complex

Electron micrographs of the ferritin antibody (rabbit) and ferritin (horse) complex have been obtained. The high iron content of the ferritin molecule (23 per cent Fe) allows its molecules to be recognized within the particles of precipitate.

Three methods of visualizing the molecular distribution have been developed: (a) small particles of the precipitated complex have been dried on to electron microscope grids and either examined directly or first shadowed with metal and then examined, (b) the precipitate has been centrifuged to a plug which was embedded and thin sections cut from it for examination, (c) the bands formed by allowing antibody and antigen to diffuse together in agar gels have been fixed, embedded and sectioned.

All methods have yielded pictures of the distribution of the ferritin within the complex which are broadly similar to what might have been expected from a somewhat irregular lattice as pictured in the Marrack-Heidelberger Lattice Theory.

The antibody molecules are not clearly defined but appear as a halo of low density enveloping the ferritin clusters. The distance, centre to centre, between the ferritin molecules is variable, but is, on the average, in the range 200–400 Å. This is greater than the ferritin-ferritin contact distance (100 Å) and is thought to mean that the ferritin molecules are bridged by antibody molecules as pictured in the Lattice Theory.

The bands produced in the gel-diffusion test contain islands of ferritin-antibody complex. When equivalent concentrations of reagents are used a single band of precipitate is formed. When excess of either antigen or antibody is used multiple bands of precipitate are formed which contain islands of ferritin antibody complex indistinguishable from those formed in the single band at equivalent concentrations, providing direct evidence for the formation of multiple bands from a single antigen.

Ferritin-ferritin contacts have been observed within the complex.

Under all the conditions of relative concentration of the two components used here, the particles of precipitated complex seem to be superficially covered with antibody which is seen as a halo about 300–400 Å thick around each cluster. This distance may correspond to the length of the antibody molecule which is deduced from other measurements to be about 300 Å.

     

Fetal damage after accidental polio vaccination of an immune mother

Irreparable damage to the anterior horn cells of the cervical and thoracic cord was found in a 20-week-old fetus whose mother was immune to poliomyelitis before conceiving but who was inadvertently given oral polio vaccine at 18 weeks gestation. Polio neutralizing antibody titres in sera, taken before and after pregnancy, were identical and were at levels normally regarded as providing protection. Unsuccessful attempts were made to isolate poliovirus from extracts of fetal brain, lung, liver and placenta. Fluorescent antibody tests were performed on various levels of the central nervous system and on the left and right extensor forearm muscles. Specific positive fluorescence to poliovirus 2 and 3 antigens was detected at dorsal spinal cord level only. One positive result was seen with Coxsackie A9 antiserum and fresh guinea-pig complement in the inflammatory cells in the right extensor forearm muscles.

This experience, as yet unexplained, underlines the importance of ensuring that women are not pregnant prior to oral polio vaccination.

     

Antibody production in mice: II. The mechanism of antigenic stimulation in the secondary immune response

To study the mechanism of antigenic stimulation in the secondary immune response, primed lymphoid cells were stimulated with antigen in various ways in vitro. The effect of antigenic stimulation was assessed by the antibody titres obtained after in vivo culture of primed cells in X-irradiated recipients.

Primed cells were much more effectively stimulated by antigen—antibody (Ag—Ab) complex than by free antigen. Primed cells could also be stimulated by spleen or lymph node cells from normal mice which had been exposed to free antigen or Ag—Ab complex in vitro or in vivo and thoroughly washed. Under these conditions, Ag—Ab complex was again much more effective than free antigen. When the cells were incubated with Ag—Ab complex, the dose of antigen bound to the cells was somewhat increased. But this increased binding of antigen could not solely account for the increase in immunogenicity.

It is suggested that the ingestion of antigen by macrophages is facilitated by the presence of antibody and that the macrophages mediate the effective immune stimulus to memory cells.

The effect of antibody in increasing the immunogenicity of antigen was lost completely when antibody was digested with pepsin. Thus, the Fc portion of antibody seemed to be important for this effect. However, it was demonstrated that antibody does not operate by becoming attached to macrophages as cytophilic antibody, and that complement is not involved in this process. The augmenting mechanism of antibody on the antigenic stimulation mediated by macrophages was discussed.

     

Development of the immune response in the foetal and newborn rhesus monkey: I. Response to bovine serum albumin

1. Foetal rhesus monkeys (at 10–16 weeks gestation) were treated in utero with bovine serum albumin (BSA) and subsequently, as neonates, were tested for immunity or tolerance.

2. Five out of six monkeys treated in utero with 100 mg BSA were tolerant to BSA at challenge.

3. Some foetuses responded to treatment with 1–8 mg BSA in adjuvant with formation of antibody.

4. One monkey treated with a total of 350 mg BSA during the first 3 weeks after birth was tolerant to BSA at challenge; two other neonatal monkeys treated with 100 mg BSA did not show tolerance.

     

Neutralizing Activity Against Clostridium difficile Toxin in the Supernatants of Cultured Colostral Cells

Human colostral specimens were obtained from 60 Japanese postpartum women within the first 3 days after delivery. Neutralizing activity against Clostridium difficile toxin was evaluated with Y1 adrenal cells in miniculture. When Y1 adrenal cells were exposed briefly to the toxin, they showed a rounding response in culture, resembling that effected by Escherichia coli enterotoxin; however, preincubation of the toxin with aqueous phase of colostrum significantly reduced its cytopathic effect on Y1 adrenal cells. Of 60 colostral specimens, 17 samples had neutralizing activity against the toxin. Cell-free supernatants of colostral cells cultured for 7 days without mitogens contained significant amounts of both immunoglobulin A (IgA) and IgM, but very small amounts of IgG. Neutralizing activity of cell-free supernatants of cultured colostral cells was evaluated as described above. Neutralizing activity against the toxin was identified in five samples of culture supernatants out of 60 colostral cell specimens. In all five cases, the aqueous phase of colostrum also had a neutralizing effect against C. difficile toxin. Neutralizing activity against the toxin found in five supernatants of cultured colostral cells was completely abolished only by anti-human IgA antibody as assessed by immune precipitation.     

Visceral larva migrans—an immunofluorescent examination of rabbit and human sera for antibodies to the ES antigens of the second stage larvae of Toxocara canis, Toxocara cati and Toxascaris leonina (Nematoda)

Precipitates were demonstrated in vitro at the orifices of the second stage larvae of Toxocara canis and T. cati when placed in sera derived from rabbits infected with these nematodes. Cross-reactions occurred between these two species.

Furthermore, these precipitates occurring at the oral, excretory pore and the anal orifices of these larvae were shown to be of a specific antibody nature by the use of the direct fluorescent antibody (Coons) staining technique.

The second stage larvae of Toxascaris leonina did not react in this way when examined in the above-mentioned experimental system, or in sera or globulins derived from rabbits infected with T. leonina.

Human sera, taken from clinically suspect cases of visceral larva migrans, were examined in this manner (q.v.). Comparable results were obtained, and it was possible to determine whether fluorescent antibodies were present, and to use this information as an aid to the clinical diagnosis of this disease.

The significance of these findings in relation to the aetiology, pathogenesis and immuno-diagnosis of visceral larva migrans is discussed.

     

Correlation between serological immune response analyzed by a new ELISA for HPV-16/18 E7 oncoprotein and clinical characteristics of cervical cancer patients

Summary. Human papillomaviruses (HPVs), particularly HPV-16/18, are linked to cervical cancer development. Full-length, recombinant HPV-16/18 E7 oncoproteins were used in a new streptavidin-biotin capture ELISA method to investigate anti-HPV E7 antibody prevalence in serum. Sera from 99 healthy women, 70 cervical cancer patients, and 30 patients with cervical pre-invasive neoplasia were analyzed. Anti-HPV-16/18 E7 positivity was found in 53% of cervical cancer patients, in 40% with cervical pre-invasive neoplasia, and in 8% of healthy women. Serum samples from 12 cervical cancer patients were obtained at different time intervals during the treatment. Eleven out of 12 showed a correspondence between HPV-E7 antibody levels (decreasing versus increasing) and the type of response (clinically complete or partial response versus progression or stable disease) at each serological evaluation. Five patients with recurrent HPV-16/18-positive cervical carcinoma were analyzed before and after vaccination with HPV-16/18 E7-pulsed autologous dendritic cells; anti-HPV-16/18 E7 positivity was found in 3 out of 5 women. In conclusion, this assay could potentially be used as an adjunctive tool to monitor the type of response to treatment and possibly to detect antibody induction in cervical cancer patients after vaccination, as a potential marker to evaluate its efficacy.     

Comparison of careHPV and Hybrid Capture 2 Assays for Detection of High-Risk Human Papillomavirus DNA in Cervical Samples from HIV-1-Infected African Women

The careHPV and HC2 assays were compared for high-risk human papillomavirus (HR-HPV) DNA detection in cervical samples from 149 HIV-1-infected African women. The HR-HPV DNA detection rates were 37.6% and 34.9% for careHPV and HC2, respectively. Agreement between the two tests was 94.6% (95% confidence interval [CI], 89.7% to 97.7%) with a kappa value of 0.88 (95% CI, 0.81 to 0.96), indicating an excellent agreement. careHPV may be considered as suitable as HC2 for cervical cancer screening among HIV-infected African women.     

Chlamydia trachomatis antigen detection in pregnancy and its verification by antibody blocking assay

Purpose: To detect the prevalence of genital infection caused by Chlamydia trachomatis in pregnant women and also to confirm the positive results using blocking antibody assay. Methods: Endocervical specimens were collected from 200 symptomatic and asymptomatic pregnant women attending the ANC OPD at M P Shah Medical College, Jamnagar. The samples were tested for presence of Chlamydia trachomatis antigen using the monoclonal antibody. Blocking antibody assay was used to further verify the positive results. Results: Out of 200 pregnant women, 38 (19%) were found positive for Chlamydia trachomatis antigen. Out of the 68 symptomatic patients, C. trachomatis antigen was detected in 26.4%. After verification of the positive samples 13.6% of the asymptomatic pregnant women were found to be harbouring the infection in their genital tract. Two (5.2%) out of the 38 positive samples, on verification with the blocking antibody assay, were found to be false positive by IDEIA,^TM thus the specificity of the IDEIA^TM being 94.8%. In patients with previous history of abortions, 27.7% were tested positive for C. trachomatis infection. Conclusions: Significant number of pregnant women shad C. trachomatis antigen in their endocervical canal, which can be easily diagnosed by this simple enzyme immuno assay having a specificity of 94.8%. Verification of positive results by antibody blocking assay can further improve the specificity of this non-culture test. Asymptomatic patients should also be screened for the infection. History of previous abortions places the patient at a higher risk for C. trachomatis infection thus such patients should be definitely tested for chlamydia infection.     

Effects of antimacrophage, antithymocyte, antilymphocyte and antispleen serum on the immune response in mice

The effect of antimacrophage serum (AMS) on antibody production was compared to that of antithymocyte (ATS) and antilymphocyte (ALS) serum. All three types of antisera inhibited antibody production to SRBC in mice. Antispleen serum was not immunosuppressive. Immunosuppression could best be demonstrated when the antisera were injected 3 days before a low dose of antigen (10⁷ or 5 × 10⁷ SRBC). None of the antisera affected secondary antibody production.

There was no correlation between the immunosuppressive potency of the antisera and their in vitro cytotoxic or in vivo lymphopenic acitivity. AMS inhibited phagocytosis, whereas ATS and ALS enhanced phagocytosis. So far we have been unable to absorb out immunosuppressive activity of the antisera but have been able to absorb out cytotoxic activity. The significance of these findings is discussed.