bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunoglobulin G (IgG) Subclass and IgE Responses in Human Paragonimiases Caused by Three Different Species
Authors: Kong Yoon , Ito Akira , Yang Hyun-Jong , Chung Young-Bae , Kasuya Shiro , Kobayashi Masashi , Liu Yue-Han , Cho Seung-Yull
Published: December 2019

In 40 cases of human paragonimiases caused byParagonimus westermani(20 cases),P. miyazakii(10 cases), andP. skrjabini(10 cases), responses of serum immunoglobulin G (IgG), IgG subclasses, and IgE were analyzed by immunoblotting with crude antigens prepared from egg, 4-week-old juvenile, and adult forms ofP. westermani. The 32- and 35-kDa proteins in the adult extracts showed specific reactions regardless of the causative species (39 of 40 cases; 98%). Sera of patients infected withP. westermaniandP. miyazakiireacted strongly with the 28-, 46-, and 94-kDa proteins of egg extracts, while those from patients infected withP. skrjabinireacted faintly. No sera from patients with other trematodiases (0 of 15 cases), cestodiases (0 of 20 cases), or lung cancer (0 of 5 cases) or from healthy controls (0 of 10 individuals) showed positive reactions. Analysis by IgG subclass revealed that IgG4 (33 of 40 cases; 83%) and IgG1 (29 of 40 cases; 73%) antibodies in the patient sera recognized the 32- and 35-kDa proteins predominantly. IgG3 reaction was found in 50% (10 of 20 cases) and 30% (3 of 10 cases) of the sera of patients infected withP. westermaniandP. miyazakii, respectively. In an IgE immunoblot, 83% (33 of 40 cases) of the sera from paragonimiasis patients reacted with the 32- and 35-kDa proteins while no sera from patients with heterologous diseases and healthy controls showed a positive reaction. Both 32- and 35-kDa proteins in adult extracts ofP. westermaniwere highly reliable for serodiagnosis of human paragonimiases.
Collection of Cervical Secretions Does Not Adversely Affect Pap Smears Taken Immediately Afterward
Authors: Hildesheim Allan , Bratti M. Concepcion , Edwards Robert P. , Schiffman Mark , Rodriguez Ana C. , Herrero Rolando , Alfaro Mario , Morera Lidia A. , Ermatinger Susan V. , Miller Barbara T. , Crowley-Nowick Peggy A.
Published: December 2019

Collection of cervical secretions for local immunological assessment requires that the secretions be collected prior to the Pap smear to avoid contamination with blood. The objective of the present study was to determine whether gentle collection of cervical secretions prior to a Pap smear collection influences the quality of the Pap smear. A total of 266 women were recruited. Half of the participants were assigned to collection of cervical secretions prior to Pap smear collection with Weck-cel sponges. The remaining half had only the Pap smear collection performed. Pap smear slides were reviewed and evaluated for quality by the Bethesda System adequacy criteria without knowledge of randomization. The proportions of limited or inadequate slides in the two study groups were compared by using the Pearson chi-square test. No significant differences were observed between the two study groups when overall Pap smear quality was evaluated (
P
= 0.29). Comparison of the two study groups with respect to individual adequacy criteria, including presence of air drying artifact, presence of obscuring blood, absence of metaplastic or endocervical cells from the transformation zone, scant cellularity, and presence of obscuring inflammatory cells, also revealed no significant differences between the two study groups. Results from the present study suggest that the collection of cervical secretions with Weck-cel sponges does not adversely impact the quality of subsequently obtained Pap smears.
Comparison of Inflammatory Events during Developing Immunoglobulin E-Mediated Late-Phase Reactions and Delayed-Hypersensitivity Reactions
Authors: Zweiman Burton , Moskovitz Anne R. , von Allmen Carolyn
Published: December 2019

To compare cellular and mediator responses in early developing late-phase skin reactions (LPR) and delayed-hypersensitivity (DH) reactions in the same subjects, responses in skin chambers overlying sites of challenge with pollen antigen andCandida albicansantigens were compared in six humans with demonstrated prominent LPR and DH responses. Histamine levels in overlying chamber fluids at 1 h were much higher at LPR than at DH sites (P= 0.002). After the next 4 h, leukocyte exudation was higher at LPR than at DH sites (P= 0.005). Most leukocytes were activated neutrophils with greater frequency of superoxide-secreting cells and released lactoferrin at LPR than at DH sites (P= 0.01 andP= 0.02, respectively). The frequency of exuding eosinophils was higher, but not significantly so (P= 0.5), at LPR sites. Although significantly more eosinophils at LPR sites were activated (P= 0.02), the levels of released eosinophilic cationic protein were not significantly higher at LPR sites (P= 0.09). The levels of interleukin-8 (IL-8), but not IL-6, were greater at LPR than at DH sites. During the first 5 h of challenge there was greater mast cell activation and subsequent exudation of activated neutrophils at sites of developing LPR than at DH sites, possibly related to greater local IL-8 levels. The frequency of activated eosinophils was also greater at LPR sites. These different initial inflammatory responses could play a role in determining expression of LPR or DH reactions.
A Euthymic Hairless Mouse Model of
Helicobacter pylori
Colonization and Adherence to Gastric Epithelial Cells In Vivo
Authors: Kimura Nobutake , Ariga Masato , Icatlo Faustino C. , Kuroki Masahiko , Ohsugi Motoyasu , Ikemori Yutaka , Umeda Kouji , Kodama Yoshikatsu
Published: December 2019

The hairless mouse strain NS:Hr/ICR was examined as a potential small animal model of
Helicobacter pylori
colonization, adherence to gastric epithelial cells in vivo, and gastritis. Among several small animals tested, NS:Hr/ICR mice proved to be the most highly susceptible to
H. pylori
infection. Challenge with clinical isolates of
H. pylori
consisting of either phenotype I or II (VacA and CagA positive and negative, respectively) resulted in colonization by mucus-resident and epithelial cell-adherent bacterial populations. Cell-adherent bacteria resisted 80 cycles of top-speed vortex washing and were recovered only by homogenization of serially washed glandular stomach tissue, indicating intimate association with the mucosal surface. Immunoperoxidase staining of paraffin sections of gastric tissue from infected mice revealed
H. pylori
antigens localized in the glandular region of the mucosa, with some colonized areas seen in the vicinity of submucosal mononuclear cell infiltration. The latter inflammatory reaction was observed as a function of the
H. pylori
phenotype (only type I induced inflammation) and the challenge dose (only those mice challenged with 10
8
CFU or higher showed the reaction). The NS:Hr/ICR strain of mice is a suitable miniature model of
H. pylori
infection and may prove useful in the quest for an efficacious mode of treatment for this common infection in humans.
Expression of Adhesion Molecules and CD28 on T Lymphocytes during Human Immunodeficiency Virus Infection
Authors: Park Suk W. , Royal Walter , Semba Richard D. , Wiegand Gordon W. , Griffin Diane E.
Published: December 2019

Adhesion molecules, which play a major role in lymphocyte circulation, have not been well characterized in human immunodeficiency virus (HIV) infection. T-lymphocyte populations, including CD3, CD4, CD28, and adhesion molecules (L selectin, LFA-1, VLA-4, and ICAM-1) were measured by flow cytometry in a cross-sectional study of 100 HIV-infected and 49 HIV-seronegative adults. HIV-infected adults had lower numbers of CD3+lymphocytes expressing L selectin (P< 0.0001) and VLA-4 (P< 0.01) and higher numbers of CD3+lymphocytes expressing LFA-1bright(P< 0.002) than did HIV-negative adults. By CD4+-lymphocyte count category (>500, 200 to 500, or <200 cells/μl), HIV-infected adults with more advanced disease had lower percentages of CD3+lymphocytes expressing L selectin and VLA-4 and higher percentages of CD3+lymphocytes expressing LFA-1. The percentages of CD3+CD28+lymphocytes and of CD3+L selectin+lymphocytes were positively correlated (Spearman coefficient = 0.86;P< 0.0001), and the percentage of CD3+CD28+lymphocytes and the CD3+LFA-1brightlymphocyte/CD3+LFA-1dimlymphocyte ratio were negatively correlated (Spearman coefficient = −0.92;P<0.00001). The results of this study suggest that HIV infection is associated with altered expression of adhesion molecules.
A Sensitive Method for Quantifying Cytomegalic Endothelial Cells in Peripheral Blood from Cytomegalovirus-Infected Patients
Authors: Kas-Deelen A. M. , Harmsen M. C. , de Maar E. F. , van Son W. J. , The T. H.
Published: December 2019

A sensitive method has been developed for the quantification of cytomegalic endothelial cells (CEC) in peripheral blood (PB) of patients with active cytomegalovirus (CMV) infections. The three subsequent key steps of this method are density centrifugation to enrich endothelial cells (EC) in the mononuclear cell (MNC) fraction, EC-specific staining, and fluorescence-activated cell sorting (FACS) of EC onto adhesion slides. The FACS method was compared with the conventional method of cytocentrifugation of the MNC fraction onto slides, followed by EC-specific staining. The main advantage of the additional steps for the isolation and quantification of CEC in PB by FACS is a 10-times-greater sensitivity than by cytocentrifugation of the MNC fraction alone. The recovery percentages of EC from whole blood were comparable for both methods. Recoveries of EC obtained after FACS were 53% ± 16.5%, (mean ± standard deviation), and recoveries of EC obtained after cytocentrifugation of the MNC fraction were 43% ± 4.3%. In patients with active CMV infection, 5 to 72 CEC were detected by FACS, equivalent to 0.8 to 9.0 CEC/ml of blood. With this method for isolation and quantification, the characterization of CEC in PB of patients with CMV-associated clinical symptoms, as well as the quantification of EC in PB of patients with pathophysiological manifestations involving endothelial damage that are different from those caused by CMV infections, can be performed.
Human Immunodeficiency Virus Type 1-Like DNA Sequences and Immunoreactive Viral Particles with Unique Association with Breast Cancer
Authors: Rakowicz-Szulczynska Eva M. , Jackson Betty , Szulczynska Adriana M. , Smith McClure
Published: December 2019

RAK antigens p120, p42, and p25 exhibit molecular and immunological similarity to the proteins encoded by human immunodeficiency virus type 1 (HIV-1) and are expressed by 95% of breast and gynecological cancer cases in women and prostate cancer cases in men. The binding of an epitope-specific anti-HIV-1 gp120 monoclonal antibody (MAb) (amino acids 308 to 322) to cancer RAK antigens has been found to be inhibited by a peptide derived from variable loop V3 of HIV-1. Breast cancer DNAs of 40 patients were PCR amplified with HIV-1 gp41-derived primers, and all of the samples were found to be positive. The DNA fragments amplified in seven blindly selected breast cancer samples were sequenced. The breast cancer DNA sequences showed at least 90% homology to the HIV-1 gene for gp41. Antisense oligonucleotides complementary to the HIV-1-like sequences inhibited reverse transcriptase activity and inhibited the growth of breast cancer cells in vitro. Viral particles detected in breast cancer cell lines were strongly immunogold labeled with the anti-HIV-1 gp120 MAb. The results obtained strongly suggest that the long-postulated breast cancer virus may, in fact, be related to HIV-1.
Diminished Adherence and/or Ingestion of VirulentMycobacterium tuberculosisby Monocyte-Derived Macrophages from Patients with Tuberculosis
Authors: Zabaleta J. , Arias M. , Maya J. R. , García L. F.
Published: December 2019

The interaction between the macrophage andMycobacterium tuberculosisis mediated by a variety of macrophage membrane-associated proteins. Complement receptors have been implicated in the adherence ofM. tuberculosisto macrophages. In the present work, the adherence and/or ingestion ofM. tuberculosisH37Rv to human monocyte-derived macrophages (MDM) from patients with tuberculosis (TB) and healthy controls was measured by microscopical examination, [3H]uracil incorporation, and CFU. The adherence and/or ingestion was enhanced by fresh serum and inhibited by heat inactivation, EDTA treatment, and anti-CR1 and anti-CR3 antibodies. Comparison of MDM from TB patients and healthy controls showed that the former exhibited a significantly decreased capacity to adhere and/or ingestM. tuberculosis, as determined by the number of CFU and3H incorporation. The expression of CR1 (CD35) and CR3 (CD11b/CD18) on MDM from TB patients and healthy controls, as determined by flow cytometry, did not show significant differences. These results suggest that the lower ingestion ofM. tuberculosisby MDM from TB patients is not due to defects in complement receptors, and therefore, there might be other molecules involved in the adherence and/or ingestion process that render MDM from TB patients ingest less mycobacteria than those from healthy controls.
Effects of Anticoagulants and Temperature on Expression of Activation Markers CD11b and HLA-DR on Human Leukocytes
Authors: Shalekoff Sharon , Page-Shipp Liesl , Tiemessen Caroline T.
Published: December 2019

A whole-blood model was used to evaluate the effects of temperature and anticoagulant on the expression of activation markers HLA-DR and CD11b on peripheral leukocytes. Venous blood, anticoagulated with either EDTA or heparin, was obtained from six healthy blood donors and 13 hospitalized patients (8 human immunodeficiency virus type 1-seropositive individuals with concurrent pulmonary tuberculosis and 5 patients with pneumonia). A preliminary evaluation was carried out with whole blood from two of the normal donors, and cells were stained immediately for HLA-DR and CD11b markers or stained after incubation at room temperature or 37°C for 18 h with or without the addition of the cytokines gamma interferon (IFN-γ), granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-γ plus GM-CSF, tumor necrosis factor beta, or interleukin-6. Of the cytokines tested, the combination of IFN-γ and GM-CSF had the most pronounced modulation of marker expression on polymorphonuclear neutrophils (PMN), in particular, HLA-DR expression, which required induction for its detection. These cytokines were therefore used in further evaluations that considered the above-mentioned effects in the presence of disease. Results indicated that the expression of activation markers on PMN and lymphocytes in whole blood are influenced by the temperature of incubation and the choice of anticoagulant and the effects noted were dependent on (i) the particular cell surface marker, (ii) the cell type being studied, and (iii) the presence or absence of disease. It is therefore recommended that ex vivo whole-blood models for evaluating phenotype or immune function be carefully evaluated for the above-mentioned effects.
Evaluation of Lipopolysaccharides and Polysaccharides of Different Epitopic Structures in the Indirect Enzyme-Linked Immunosorbent Assay for Diagnosis of Brucellosis in Small Ruminants and Cattle
Authors: Alonso-Urmeneta B. , Marín C. , Aragón V. , Blasco J. M. , Díaz R. , Moriyón I.
Published: December 2019

Brucella abortus
and
Brucella melitensis
have surface lipopolysaccharides and polysaccharides carrying
B. melitensis
-type (M) and
B. abortus
-type (A) epitopes as well as common (C) epitopes present in all smooth
Brucella
biotypes. Crude lipopolysaccharides, hydrolytic O polysaccharides, and native hapten polysaccharides of MC or AC specificity were evaluated in indirect enzyme-linked immunosorbent assays with polyclonal, monoclonal, or protein G conjugates by using sera from cattle, sheep, and goats infected with AC, MC, or AMC
Brucella
biotypes. Regardless of the antigen, the levels of antibodies were lower in goats than in sheep and highest in cattle. The diagnostic performance of the assay was not affected by the absence of lipid A-core epitopes, the presence of contaminating outer membrane proteins, the AC or MC epitopic structure of the absorbed antigen, or the conjugate used. Moreover, with sera from cattle vaccinated with
B. abortus
S19 (AC) or from sheep and goats vaccinated with
B. melitensis
Rev 1 (MC), AC and MC antigens showed similar levels of reactivity. The results show that antibodies to the C epitopes largely dominate in infection, and this is consistent with the existence of multiple overlapping C epitopes (V. Weynants, D. Gilson, A. Cloeckaert, A. Tibor, P. A. Denoel, F. Godfroid, J. N. Limet, and J.-J. Letesson, Infect. Immun. 65:1939–1943, 1997) rather than with one or two C epitopes. It is concluded that, by adaptation to the corresponding antibody levels, brucellosis in cattle, sheep, and goats can be diagnosed by immunosorbent assay with a single combination of conjugate and antigen.
Serologic Evidence of a Natural Infection of White-Tailed Deer with the Agent of Human Granulocytic Ehrlichiosis in Wisconsin and Maryland
Authors: Walls Jennifer J. , Asanovich Kristin M. , Bakken Johan S. , Dumler J. Stephen
Published: December 2019

White-tailed deer participate in the maintenance of the
Ixodes
tick life cycle and are reservoirs for some tick-borne infectious agents. Deer may be useful sentinels for tick-transmitted agents, such as ehrlichiae. In order to determine whether white-tailed deer are markers of natural transmission or are reservoirs for the human granulocytic ehrlichiosis (HGE) agent, we performed indirect immunofluorescent-antibody (IFA) tests and immunoblotting with the HGE agent and
Ehrlichia chaffeensis
on sera from 43 and 294 deer captured in northwest Wisconsin during 1994 and 1995, respectively, and 12 deer from southern Maryland. According to IFA testing, 47% of 1994 Wisconsin sera, 60% of 1995 Wisconsin sera, and 25% of Maryland sera contained HGE agent antibodies. All IFA-positive deer sera tested reacted with the 44-kDa band which is unique to the
Ehrlichia phagocytophila
group. Serologic reactions to
E. chaffeensis
were detected by IFA testing in 15 of 337 (4%) Wisconsin deer and in 10 of 12 (83%) Maryland deer, while 60 and 80% of
E. chaffeensis
IFA-positive Wisconsin and Maryland deer sera, respectively, reacted with the
E. chaffeensis
28- to 29-kDa antigens by immunoblotting. A total of 4% of deer from Wisconsin and 25% of deer from Maryland were found by IFA testing to have antibodies to both the HGE agent and
E. chaffeensis
; 75% of these were confirmed to contain
E. chaffeensis
antibodies by immunoblotting. These results suggest that white-tailed deer in diverse geographical regions of the United States are naturally infected with the HGE agent,
E. chaffeensis
, or both and that these animals, and potentially humans, are exposed to infected ticks at a high frequency in nature.
Effect of Shipment, Storage, Anticoagulant, and Cell Separation on Lymphocyte Proliferation Assays for Human Immunodeficiency Virus-Infected Patients
Authors: Weinberg Adriana , Betensky Rebecca A. , Zhang Li , Ray Graham
Published: December 2019

Lymphocyte proliferation assays (LPA), which can provide important information regarding the immune reconstitution of human immunodeficiency virus (HIV)-infected patients on highly active antiretroviral therapy, frequently involve shipment of specimens to central laboratories. In this study, we examine the effect of stimulant, anticoagulant, cell separation, storage, and transportation on LPA results. LPA responses of whole blood and separated peripheral blood mononuclear cells (PBMC) to different stimulants (cytomegalovirus, varicella-zoster virus, candida and tetanus toxoid antigens, and phytohemagglutinin) were measured using fresh specimens shipped overnight and frozen specimens collected in heparin, acid citrate dextrose (ACD), and citrate cell preparation tubes (CPT) from 12 HIV-infected patients and uninfected controls. Odds ratios for positive LPA responses were significantly higher in separated PBMC than in whole blood from ACD- and heparin-anticoagulated samples obtained from HIV-infected patients and from ACD-anticoagulated samples from uninfected controls. On separated PBMC, positive responses were significantly more frequent in fresh samples compared with overnight transportation for all antigens and compared with cryopreservation for the candida and tetanus antigens. In addition, viral antigen LPA responses were better preserved in frozen PBMC compared with specimens shipped overnight. CPT tubes yielded significantly more positive LPA results for all antigens, irrespective of the HIV patient status compared with ACD, but only for the candida and tetanus antigens and only in HIV-negative controls compared with heparin. Although HIV-infected patients had a significantly lower number of positive antigen-driven LPA responses compared with uninfected controls, most of the specimen processing variables had similar effects on HIV-positive and -negative samples. We conclude that LPA should be performed on site, whenever feasible, by using separated PBMC from fresh blood samples collected in either heparin or ACD. However, if on-site testing is not available, optimal transportation conditions should be established for specific antigens.
Serological Assessment of the Early Response to Eradication Therapy Using an Immunodominant Outer Membrane Protein of
Helicobacter pylori
Authors: Nishizono Akira , Gotoh Takayuki , Fujioka Toshio , Murakami Kazunari , Kubota Toshihiro , Nasu Masaru , Watanabe Makoto , Mifune Kumato
Published: December 2019

Eradication of
Helicobacter pylori
infection cures gastritis and prevents recurrence of peptic ulcers. Endoscopy is usually used to evaluate the effectiveness of eradication therapy. We designed a new noninvasive assay system for the early evaluation of eradication of
H. pylori
infection in which a crude
H. pylori
outer membrane protein preparation (HPOmp) is used as an antigen, and we determined the sensitivity and specificity of the serological assay system. Immunoblot analysis showed that anti-HPOmp antibodies reacted to a protein with a molecular mass of approximately 29 kDa. In those patients who responded to therapy, the anti-HPOmp immunoglobulin G (IgG) titers measured by enzyme-linked immunosorbent assay (ELISA) at 1 month after the end of therapy were significantly lower than those before treatment (34.8% reduction;
P
< 0.001), and the posttreatment reduction in the antibody titer was significantly greater than that of the titer measured with a commercially available anti-
H. pylori
IgG ELISA (34.8% versus 16.1%;
P
< 0.001). When a 25% reduction of anti-HPOmp IgG titer at 1 month after the end of treatment was taken as the cutoff value for
H. pylori
eradication, the sensitivity and specificity of our new assay were 75% (51 of 68 treatment responders) and 96% (22 of 23 nonresponders), respectively. Our results indicate that the novel serological test with HPOmp might be a clinically useful tool for assessment of eradication of
H. pylori
.
Aging and the Immune Response to Tetanus Toxoid: Diminished Frequency and Level of Cellular Immune Reactivity to Antigenic Stimulation
Authors: Schatz Desmond , Ellis Tamir , Ottendorfer Eric , Jodoin Eric , Barrett Douglas , Atkinson Mark
Published: December 2019

The period of efficacious immune reactivity afforded by tetanus immunization and the need for continuing some forms of tetanus vaccination programs have been the subjects of recent debates. Our studies demonstrate that the level of antitetanus immunity based on immunological memory (i.e., cellular immune responsiveness) varies dramatically as a function of age, with older individuals constituting a population which is increasingly susceptible to tetanus infection.
Development and Applications of a Bovine Coronavirus Antigen Detection Enzyme-Linked Immunosorbent Assay
Authors: Schoenthaler S. L. , Kapil S.
Published: December 2019

We developed a monoclonal antibody-based, antigen capture sandwich enzyme-linked immunosorbent assay (ELISA) for bovine coronavirus. We compared the ELISA with electron microscopy and the hemagglutination test and found a close correlation between them. The sensitivity of the ELISA was 104bovine coronavirus particles per ml of 10% fecal suspension. Compared with electron microscopy, bovine coronavirus ELISA had 96% specificity.
Humoral Immune Response to Human Cytomegalovirus in Patients Undergoing Percutaneous Transluminal Coronary Angioplasty
Authors: Tiran Andreas , Tio Rene A. , Oostenveld Esther , Harmsen Martin C. , Tiran Beate , Den Heijer Peter , Monnink Stefan H. J. , Wilders-Truschnig Martie M. , The T. Hauw
Published: December 2019

Possible causal relations between prior human cytomegalovirus (HCMV) infection and atherosclerosis and between HCMV reactivation and restenosis after coronary angioplasty have been suggested. We investigated patterns of antibodies directed to HCMV in 112 patients undergoing percutaneous transluminal coronary angioplasty (PTCA) and in a group of sex- and age-matched controls (blood donors without evidence of atherosclerosis). Levels of antibodies to HCMV were measured by enzyme-linked immunosorbent assay (ELISA) of serum samples drawn before and 5 weeks after PTCA. To further differentiate the humoral immune response, we specifically tested antibody reactivity towards four single HCMV proteins (IE2, p52, pp150, and pp65) by recombinant ELISAs. We found that 73% of PTCA patients and 69% of sex- and age-matched controls were seropositive for HCMV (odds ratio, 1.2 [not significant]). The corresponding odds ratios for matched pairs ranged in the recombinant ELISAs from 1.2 to 1.4. Patients had more often high titers of anti-HCMV antibodies (11 versus 4%; odds ratio = 3.3 [0.9 to 15.2];
P
= 0.052) and high titers of anti-pp150 antibodies (13 versus 4%; odds ratio = 6.0 [1.3 to 38.8];
P
= 0.008). Anti-HCMV immunoglobulin M antibodies were not detected in any patient. There was no evidence of acute HCMV reactivation after PTCA, since the titers of antibodies to the investigated recombinant proteins did not increase at 5 weeks after PTCA. Our results show a limited association between prior HCMV infection and coronary artery disease. We infer that positive anti-HCMV titers are not a major risk factor at the time of disease manifestation. However, this study cannot rule out a possible role of HCMV at earlier stages of the atherosclerotic process. Recombinant ELISAs provide a valuable tool for investigating the antiviral immune response.
The Cured Immune Phenotype Achieved by Treatment of Visceral Leishmaniasis in the BALB/c Mouse with a Nonionic Surfactant Vesicular Formulation of Sodium Stibogluconate Does Not Protect against Reinfection
Authors: Carter K. C. , Baillie A. J. , Mullen A. B.
Published: December 2019

Single-dose treatment with sodium stibogluconate solution (SSG) and treatment with a nonionic surfactant vesicular formulation of sodium stibogluconate (SSG-NIV) were compared for the ability to protect BALB/c mice against infection withLeishmania donovani.Prophylactic treatment with SSG-NIV protected against infection, although its effects were time and organ dependent; protection was not obtained with SSG. Protection against reinfection withL. donovaniwas observed only in mice cured by treatment with SSG-NIV. However, this protective effect was probably due to the presence of residual drug rather than an immune effect, since prophylactic SSG-NIV treatment gave similar results. Transfer of enriched spleen T-cell populations fromL. donovani-infected mice or from infected SSG-NIV-treated mice gave no protection againstL. donovaniinfection in the recipients. T cells from infected mice, but not from infected SSG-NIV-treated mice, were infectious to recipients. SSG-NIV treatment was equally effective against visceral leishmaniasis in immunocompetent and SCID mice, whereas SSG treatment was less effective in the latter. The results of this study suggest that the high antileishmanial activity of SSG-NIV is due to favorable modification of SSG delivery and does not require a fully functional immune response. Cure of visceral leishmaniasis by SSG-NIV treatment in the BALB/c mouse did not protect against reinfection.
Immune Determinants of Organism and Outcome in Febrile Hospitalized Thai Patients with Bloodstream Infections
Authors: Jason Janine , Archibald Lennox , McDonald L. Clifford , Hart W. Michael , Rheanppumikankit Sunthorn , Tansuphwaswadikul Somsit , Byrd Martha G. , Larned Joshua , Han Alison , Green Timothy A. , Jarvis William R.
Published: December 2019

Opportunistic infections (OI) and the human immunodeficiency virus (HIV) cause significant morbidity and mortality in developing countries. Immune cell and cytokine profiles may be related to the type and course of OI and to the OI-HIV interaction. Examining cell-specific cytokine production ex vivo has only recently become feasible. In Thailand, 53 febrile, hospitalized adults were enrolled in a study of the immune correlates of bloodstream infections (BSI). On site, blood cells were stimulated ex vivo. Cell-surface antigens and eight intracellular cytokines were subsequently analyzed using flow cytometry to determine associations with mortality and the organism causing the BSI. By logistic regression analysis, the percentage of CD3+CD16/56+cells making tumor necrosis factor alpha (TNF-α) (P= 0.033) and the percentage of CD3−CD16/56+cells (NK) (P= 0.032) were related to HIV positivity. Lymph node enlargement with HIV infection and the percentage of CD3+CD16/56+making TNF-α were predictive of death. A lower percentage of CD3+CD8+lymphocytes making interleukin-8 (IL-8) (P= 0.005), fewer monocytes expressing CD14 (P= 0.009), and the percentage of CD3+CD8+cells producing gamma interferon (P= 0.011) were associated with blood culture positivity and the causative organism. For every one point decrease in the percentage of CD3+CD8+cells making IL-8, the likelihood of a positive culture increased 23%; for every one point decrease in the percentage of monocytes expressing CD14, the likelihood of a positive culture increased by 5%. Only a few immune cell types and three of their related cytokines were significantly associated with HIV disease outcome or the BSI organism. These cell types did not include CD3+CD8−cells (a surrogate for CD4+cells), nor did they involve cytokines associated with a type I to type II cytokine shift, which might occur with advancing HIV infection. These associations support the premise that CD8+and CD16/56+lymphocytes play significant roles in HIV and type I infections.
Quantitative Analysis of T-Cell Receptor β Variable-Gene Usage in Cutaneous Late-Phase Reactions: Implications for T-Lymphocyte Recruitment in Cutaneous Inflammation
Authors: Lessin Stuart R. , Benoit Bernice M. , Li Guoqing , Moskovitz Ann , Zweiman Burton
Published: December 2019

To determine if functionally distinct T-lymphocyte (T cell) subsets accumulate in late-phase immunoglobulin E-mediated reactions (LPR), we quantitatively analyzed the immunophenotype and the T-cell receptor β variable-gene (Vβ) repertoire of T cells in cutaneous LPR. Peripheral blood and skin biopsies were obtained 6 or 24 h after sensitive subjects were challenged with intradermal injections of grass pollen allergen (Ag) and control (C) solution. The frequency of cells expressing CD3, CD4, CD8, CD45RO, and CD25/mm2was determined by immunohistochemistry in nine subjects. Vβ usage was assessed by reverse transcription-PCR in five of nine subjects. A significantly greater frequency of CD3+and CD45RO+(memory) T cells was detected in Ag sites than in C sites at 24 h after challenge but not at 6 h. The frequency of activated (CD25+) and helper (CD4+) T cells appeared to be increased in Ag sites as well, though not significantly. Vβ6 was the most commonly expressed Vβ detected in Ag sites, but it was also detected in accompanying C sites. Vβ2 was the most commonly expressed Vβ detected in C sites. Sequence analysis in one case revealed Vβ expression in a 6-h Ag site to be essentially polyclonal. Our findings suggest that memory T cells with Vβ expression similar to that in normal skin accumulate in developing cutaneous LPR. The limited usage of Vβ suggests a preferential recruitment or retention of reactive T cells from an endogenous subset of skin-homing T cells with its own skewed Vβ repertoire.
Variables That Affect Assays for Plasma Cytokines and Soluble Activation Markers
Authors: Aziz Najib , Nishanian Parunag , Mitsuyasu Ronald , Detels Roger , Fahey John L.
Published: December 2019

Cytokines and soluble immune activation markers that reflect cytokine activities in vivo are increasingly being measured in plasma, serum, and other body fluids. They provide useful diagnostic and prognostic information as well as insight into disease pathogenesis. Assays of neopterin, β2-microglobulin, soluble interleukin-2 receptor, and soluble tumor necrosis factor receptor type II as well as of the cytokines tumor necrosis factor alpha and gamma interferon (IFN-γ) were evaluated by using serum and plasma samples of human immunodeficiency virus (HIV)-positive and HIV-negative subjects. Many factors were found to influence the outcomes of these assays. Substantial differences in apparent levels of analytes were frequently found when enzyme-linked immunosorbent assay (ELISA) kits from different manufacturers were used. In some cases, differences were found in the standards provided by separate manufacturers. Furthermore, the analytic results from different lots of ELISA kits supplied by single manufacturers differed by as much as 50%. The need for uniformity in the standards for quantitative assays was clearly illustrated. International reference standards are available for cytokines but not for soluble cytokine receptors or soluble activation markers. Marker levels in serum or in plasma were similar except those for IFN-γ. Most of the analytes were stable under several storage conditions. Thus, batch testing of frozen stored samples is feasible. The findings indicate that for longitudinal studies, the levels of cytokines and immune activation markers in plasma or serum should be measured by using preverified reagents from one manufacturer. The quality of laboratory performance can have an impact on clinical relevance. Proficiency testing and external quality assurance programs can help to develop the needed consensus.
Plasma-Soluble CD30 in Childhood Tuberculosis: Effects of Disease Severity, Nutritional Status, and Vitamin A Therapy
Authors: Hanekom W. A. , Hussey G. D. , Hughes E. J. , Potgieter S. , Yogev R. , Check I. J.
Published: December 2019

Plasma-soluble CD30 (sCD30) is the result of proteolytic splicing from the membrane-bound form of CD30, a putative marker of type 2 cytokine-producing cells. We measured sCD30 levels in children with tuberculosis, a disease characterized by prominent type 1 lymphocyte cytokine responses. We postulated that disease severity and nutritional status would alter cytokine responses and therefore sCD30 levels. Samples from South African children enrolled prospectively at the time of diagnosis of tuberculosis were analyzed. (Patients were originally enrolled in a randomized, double-blind placebo-controlled study of the effects of oral vitamin A supplementation on prognosis of tuberculosis.) Plasma samples collected at the time of diagnosis and 6 and 12 weeks later (during antituberculosis therapy) were analyzed. sCD30 levels were measured by enzyme immunoassay. The 91 children included in the study demonstrated high levels of sCD30 at diagnosis (median, 98 U/liter; range, 11 to 1,569 U/liter). Although there was a trend toward higher sCD30 levels in more severe disease (e.g., culture-positive disease or miliary disease), this was not statistically significant. Significantly higher sCD30 levels were demonstrated in the presence of nutritional compromise: the sCD30 level was higher in patients with a weight below the third percentile for age, in those with clinical signs of kwashiorkor, and in those with a low hemoglobin content. There was minimal change in the sCD30 level after 12 weeks of therapy, even though patients improved clinically. However, changes in sCD30 after 12 weeks differed significantly when 46 patients (51%) who received vitamin A were compared with those who had received a placebo. Vitamin A-supplemented children demonstrated a mean (± standard error of the mean) decrease in sCD30 by a factor of 0.99 ± 0.02 over 12 weeks, whereas a factor increase of 1.05 ± 0.02 was demonstrated in the placebo group (P= 0.02). We conclude that children with tuberculosis had high sCD30 levels, which may reflect the presence of a type 2 cytokine response. Nutritional compromise was associated with higher sCD30 levels. Vitamin A therapy resulted in modulation of sCD30 levels over time.
Identification of
Acinetobacter baumannii
Strains with Monoclonal Antibodies against the O Antigens of Their Lipopolysaccharides
Authors: Pantophlet Ralph , Brade Lore , Brade Helmut
Published: December 2019

Despite the emergence of
Acinetobacter baumannii
strains as nosocomial pathogens, simple methods for their phenotypic identification are still unavailable. Murine monoclonal antibodies specific for the O-polysaccharide moiety of the lipopolysaccharide (LPS) of two
A. baumannii
strains were obtained after immunization with heat-killed bacteria. The monoclonal antibodies were characterized by enzyme immunoassay and by Western and dot blot analyses and were investigated for their potential use for the identification of
A. baumannii
strains. The antibodies reacted with 46 of the 80
A. baumannii
clinical isolates that were investigated, and reactivity was observed with 11 of 14 strains which were isolated during outbreaks in different northwestern European cities; no reactivity was observed with
Acinetobacter
strains of other genomic species, including the closely related genomic species 1 (
Acinetobacter calcoaceticus
), 3, and 13 sensu Tjernberg and Ursing, or with other gram-negative bacterial strains. The results show that O-antigen-specific monoclonal antibodies such as the ones described are convenient reagents which can be used to identify
Acinetobacter
strains in clinical and research laboratories.
Detection of Cryptdin in Mouse Skin
Authors: Shirafuji Yoshinori , Oono Takashi , Kanzaki Hiroko , Hirakawa Satoshi , Arata Jirô
Published: December 2019

Defensins are widely distributed and broad-spectrum antimicrobial peptides with activities against bacteria, fungi, and enveloped viruses. Defensins have been isolated from granules of neutrophils from humans, rabbits, rats, and guinea pigs. They have also been found in lung macrophages as well as in Paneth cells of the human, rabbit, and mouse small intestine. The human β-defensin-2 was recently isolated from human skin. In this study, we detected the expression of mRNA for the defensin cryptdin in BALB/c mouse skin by means of reverse transcriptase PCR amplification. Expression was also detected in dispase-separated epidermis and cultured keratinocytes, but expression was not detected in fibroblasts. The expression of cryptdin mRNA was found to begin on embryonic day 17.5. As determined with specific primers, the cDNA sequence cloned from the skin was found to be identical to that previously reported for cryptdin-5. cDNA derived from cultured keratinocytes demonstrated the sequences of the cryptdin-6 and cryptdin-1 isoforms. In situ hybridization analysis showed that the mRNA of cryptdin was expressed in the suprabasal keratinocytes of the skin in embryonic and neonatal days and then shifted to the hair bulbs in the skin of adult mice.
Immunohematological Reference Ranges for Adult Ethiopians
Authors: Tsegaye Aster , Messele Tsehaynesh , Tilahun Tesfaye , Hailu Ermias , Sahlu Tefera , Doorly Ronan , Fontanet Arnaud L. , Rinke de Wit Tobias F.
Published: December 2019

A cross-sectional survey was carried out with 485 healthy working adult Ethiopians who are participating in a cohort study on the progression of human immunodeficiency virus type 1 (HIV-1) infection to establish hematological reference ranges for adult HIV-negative Ethiopians. In addition, enumeration of absolute numbers and percentages of leukocyte subsets was performed for 142 randomly selected HIV-negative individuals. Immunological results were compared to those of 1,356 healthy HIV-negative Dutch blood donor controls. Immunohematological mean values, medians, and 95th percentile reference ranges were established. Mean values were as follows: leukocyte (WBC) counts, 6.1 × 10
9
/liter (both genders); erythrocyte counts, 5.1 × 10
12
/liter (males) and 4.5 × 10
12
/liter (females); hemoglobin, 16.1 (male) and 14.3 (female) g/dl; hematocrit, 48.3% (male) and 42.0% (female); platelets, 205 × 10
9
/liter (both genders); monocytes, 343/μl; granulocytes, 3,057/μl; lymphocytes, 1,857/μl; CD4 T cells, 775/μl; CD8 T cells, 747/μl; CD4/CD8 T-cell ratio, 1.2; T cells, 1,555/μl; B cells, 191/μl; and NK cells, 250/μl. The major conclusions follow. (i) The WBC and platelet values of healthy HIV-negative Ethiopians are lower than the adopted reference values of Ethiopia. (ii) The absolute CD4 T-cell counts of healthy HIV-negative Ethiopians are considerably lower than those of the Dutch controls, while the opposite is true for the absolute CD8 T-cell counts. This results in a significantly reduced CD4/CD8 T-cell ratio for healthy Ethiopians, compared to the ratio for Dutch controls.
Expression of Fas (CD95/APO-1) Ligand by Human Breast Cancers: Significance for Tumor Immune Privilege
Authors: O’Connell Joe , Bennett Michael W. , O’Sullivan Gerald C. , O’Callaghan Jim , Collins J. Kevin , Shanahan Fergus
Published: December 2019

Breast cancers have been shown to elicit tumor-specific immune responses. As in other types of cancer, the antitumor immune response fails to contain breast tumor growth, and a reduction in both the quantity and cytotoxic effectiveness of tumor-infiltrating lymphocytes (TILs) is associated with a poorer prognosis. Fas ligand (FasL) induces apoptotic death of activated lymphocytes that express its cell surface receptor, FasR (CD95/APO-1). FasL-mediated apoptosis of activated lymphocytes contributes to normal immune downregulation through its roles in tolerance acquisition, immune response termination, and maintenance of immune privilege in the eye, testis, and fetus. In this report, we demonstrate that breast carcinomas express FasL. Using in situ hybridization and immunohistochemistry, we show that breast tumors constitutively express FasL at both the mRNA and protein levels, respectively. FasL expression is prevalent in breast cancer: 100% of breast tumors (17 of 17) were found to express FasL, and expression occurred over more than 50% of the tumor area in all cases. By immunohistochemistry, FasR was found to be coexpressed with FasL throughout large areas of all the breast tumors. This suggests that the tumor cells had acquired intracellular defects in FasL-mediated apoptotic signaling. FasL and FasR expression were independent of tumor type or infiltrative capacity. FasL expressed by tumor cells has previously been shown to kill Fas-sensitive lymphoid cells in vitro and has been associated with apoptosis of TILs in vivo. We conclude that mammary carcinomas express FasL in vivo as a potential inhibitor of the antitumor immune response.
Detection of Anti-VacA Antibody Responses in Serum and Gastric Juice Samples Using Type s1/m1 and s2/m2
Helicobacter pylori
VacA Antigens
Authors: Perez-Perez Guillermo I. , Peek Richard M. , Atherton John C. , Blaser Martin J. , Cover Timothy L.
Published: December 2019

Several different families of vacuolating toxin (
vacA
) alleles are present in
Helicobacter pylori
, and they encode products with differing functional activities.
H. pylori
strains containing certain types of
vacA
alleles have been associated with an increased risk for peptic ulcer disease. In this study, we tested serum samples and gastric juice from 19
H. pylori
-negative and 39
H. pylori
-positive patients for enzyme-linked immunosorbent assay reactivity with two different types of VacA antigens (types s1/m1 and s2/m2), which were purified from
H. pylori
60190 and 86-338, respectively. Both antigens were recognized better by serum immunoglobulin G (IgG) from
H. pylori
-positive persons than by serum IgG from
H. pylori
-negative persons (
P
< 0.01). The s1/m1 VacA antigen was better recognized by sera from patients carrying
vacA
type s1/m1 strains than by sera from patients carrying
vacA
type s2/m2 or s1/m2 strains (
P
< 0.01). Conversely, the s2/m2 VacA antigen was better recognized by sera from patients carrying type s2/m2 or s1/m2 strains (
P
= 0.03). Serum IgG anti-VacA antibodies were present more frequently in patients carrying type s1/m1 strains than in other
H. pylori
-positive patients (
P
= 0.0002). In addition, the highest levels of IgA anti-VacA antibodies were detected in the gastric juice of patients carrying type s1/m1 strains. These data indicate that different VacA isoforms have distinct antigenic properties and that multiple forms of VacA elicit antibody responses in
H. pylori
-positive humans.
Pneumococcal Capsular Polysaccharide Preparations May Contain Non-C-Polysaccharide Contaminants That Are Immunogenic
Authors: Yu Xinhong , Sun Yan , Frasch Carl , Concepcion Nelydia , Nahm Moon H.
Published: December 2019

We measured the capacity to opsonize
Streptococcus pneumoniae
serotype 6B and estimated the concentration of immunoglobulin G anti-6B capsular polysaccharide (PS) antibodies in 25 pre- and postimmune sera from adults immunized with a pneumococcal PS vaccine. We first studied two postvaccination serum samples displaying less opsonophagocytic capacity than expected. The majority of anti-6B antibodies in the two samples reacted with the capsular PSs of several unrelated serotypes (2, 4, 9V, 19F, and 23F) and with the lysate of noncapsulated
S. pneumoniae
bacteria but not with C-PS. The non-type-specific antibodies accounted for at least one-half of anti-6B antibodies in 40% of prevaccination sera and 10% of postvaccination sera from adults. The non-type-specific antibodies could be demonstrated in the enzyme-linked immunosorbent assays (ELISAs) for pneumococcal antibodies to other serotypes (4, 9V, 18C, 19F, and 23F). The nonspecific antibodies appear to bind a contaminant(s) in the current preparations of capsular PS. ELISA for antibodies to pneumococcal capsules may not be serotype specific for some samples.
Effects of Amphetamine on Development of Oral Candidiasis in Rats
Authors: Freire-Garabal M. , Núñez M. J. , Balboa J. , Rodríguez-Cobo A. , López-Paz J. M. , Rey-Méndez M. , Suárez-Quintanilla J. A. , Millán J. C. , Mayán J. M.
Published: December 2019

Experiments were conducted to evaluate the effects of amphetamine (0.4 mg/kg of body weight/day) on the development of oral candidiasis in Sprague-Dawley rats. Animals were submitted to surgical hyposalivation in order to facilitate the establishment and persistence of
Candida albicans
infection. Treatment with drugs (placebo or amphetamine) was initiated 7 days before
C. albicans
inoculation and lasted until the end of the experiments, day 15 postinoculation. Establishment of
C. albicans
infection was evaluated by swabbing the inoculated oral cavity with a sterile cotton applicator on days 2 and 15 after inoculation, followed by plating on YEPD (yeast extract-peptone-dextrose) agar. Tissue injury was determined by the quantification of the number and type (normal or abnormal) of papillae on the dorsal tongue per microscopic field. A semiquantitative scale was devised to assess the degree of colonization of the epithelium by fungal hyphae. Our results show that amphetamine exacerbates
C. albicans
infection of the tongues of rats. Significant increases in
Candida
counts, the percentage of the tongue’s surface covered with clinical lesions, the percentage of abnormal papillae, and the colonization of the epithelium by fungal hyphae were found in amphetamine-treated rats compared to those found in the rats injected with a placebo. The last two parameters increased in rats treated with the placebo compared to the parameters of the untreated control rats.