SERO-REACTIVITY PROTOTYPE OF SECRETED PROTEINS OF NATIVE ‘S 5’ VACCINE STRAIN TO HUMAN SERA POSITIVE FOR Mycobacterium paratuberculosis INFECTION

Our long-term goal is to develop an efficient acellular vaccine against paratuberculosis based on protein antigen(s). A prerequisite to achieve this goal is to analyze and characterize Mycobacterium paratuberculosis (Mpt) secreted and cellular proteins eliciting a protective immune response. In the context of this general objective, we proposed to identify, clone, produce, and characterize: the Mpt 85B antigen and other Mpt immunoreactive secreted proteins, the Mpt L7/L12 ribosomal protein and other immunoreactive cellular proteins, Mpt protein determinants involved in invasion of epithelial cells, and Mpt protein antigens specifically expressed in macrophages. Paratuberculosis is still a very serious problem in Israel and in the USA. In the USA, a recent survey evaluated that 21.6% of the dairy herd were infected with Mpt resulting in 200-250 million dollars in annual losses. Very little is known on the virulence factors and on protective antigens of Mpt. At present, the only means of controlling this disease are culling or vaccination. The current vaccines do not allow a clear differentiation between infected and vaccinated animals. Our long-term goal is to develop an efficient acellular paratuberculosis vaccine based on Mpt protein antigen(s) compatible with diagnostic tests. To achieve this goal it is necessary to analyze and characterize secreted and cellular proteins candidate for such a vaccine. Representative Mpt libraries (shuttle plasmid and phage) were constructed and used to study Mpt genes and gene products described below and will be made available to other research groups. In addition, two approaches were performed which did not yield the expected results. Mav or Mpt DNA genes that confer upon Msg or E. coli the ability to invade and/or survive within HEp-2 cells were not identified. Likewise, we were unable to characterize the 34-39 kDa induced secreted proteins induced by stress factors due to technical difficulties inherent to the complexity of the media needed to support substantial M. pt growth. We identified, isolated, sequenced five Mpt proteins and expressed four of them as recombinant proteins that allowed the study of their immunological properties in sensitized mice. The AphC protein, found to be up regulated by low iron environment, and the SOD protein are both involved in protecting mycobacteria against damage and killing by reactive oxygen (Sod) and nitrogen (AhpC) intermediates, the main bactericidal mechanisms of phagocytic cells. SOD and L7/L12 ribosomal proteins are structural proteins constitutively expressed. 85B and CFP20 are both secreted proteins. SOD, L7/L12, 85B and CFP20 were shown to induce a Th1 response in immunized mice whereas AphC was shown by others to have a similar activity. These proteins did not interfere with the DTH reaction of naturally infected cows. Cellular immunity provides protection in mycobacterial infections, therefore molecules inducing cellular immunity and preferentially a Th1 pathway will be the best candidate for the development of an acellular vaccine. The proteins characterized in this grant that induce a cell-mediated immunity and seem compatible with diagnostic tests, are good candidates for the construction of a future acellular vaccine.

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Characterization of a Nonspecific Inhibitor Found in Human Sera Raised Against the 2006/07 Influenza Vaccine Strain A/Hiroshima/52/2005 (H3N2) Virus

Kansenshogaku zasshi ◽

10.11150/kansenshogakuzasshi.86.400 ◽

2012 ◽

Vol 86 (4) ◽

pp. 400-404

Author(s):

Akiko MAEDA ◽

Saeko MORIKAWA ◽

Tetsuo KASE ◽

Sin IRIE ◽

Yoshio HIROTA

Keyword(s):

Influenza Vaccine ◽

Vaccine Strain ◽

H3n2 Virus ◽

Human Sera

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A library of cell-surface and secreted proteins fromSchistosoma mansoniidentifies early serological markers of infection

10.1101/593616 ◽

2019 ◽

Cited By ~ 1

Author(s):

Cécile Crosnier ◽

Anna V. Protasio ◽

Gabriel Rinaldi ◽

Shona Wilson ◽

Matthew Berriman ◽

...

Keyword(s):

Recombinant Protein ◽

Cell Surface ◽

Mammalian Cells ◽

Treatment Strategies ◽

Secreted Proteins ◽

Serological Markers ◽

Low Transmission ◽

Protein Library ◽

Wide Range ◽

Human Sera

AbstractSchistosomiasis is a major global health problem caused by blood-dwelling parasitic worms and is currently treated by the mass administration of the drug praziquantel. Appropriate drug treatment strategies are informed by diagnostics that establish the prevalence and intensity of infection, which, in regions of low transmission should be highly sensitive. To identify sensitive new serological markers ofSchistosoma mansoniinfections, we have compiled a recombinant protein library of 115 parasite cell surface and secreted proteins expressed in mammalian cells. The vast majority of them were shown to be immunoreactive and to contain heat-labile conformational epitopes when tested against pooled human sera from endemic regions. After probing the library against a time series of sera samples from experimental infections in mice, we identified several markers of infection, the majority of which belong to the saposin-domain-containing and cathepsin families of proteins. These new markers will be a valuable tool to detect ongoing and previousS. mansoniinfections, including in regions of low transmission. We envisage that this new recombinant protein resource will be used in a wide range of cellular and molecular assays to further our understanding ofSchistosomabiology.

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Immunological Evidence for Prothrombin in Human Platelets

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661534 ◽

1986 ◽

Vol 55 (02) ◽

pp. 268-270

Author(s):

R J Alexander

Keyword(s):

Electrophoretic Mobility ◽

Platelet Activation ◽

Double Diffusion ◽

Human Platelets ◽

Secreted Proteins ◽

Platelet Surface ◽

Activated Platelets ◽

Similar Protein ◽

Diffusion Assay ◽

Supernatant Solution

SummaryAn attempt was made to isolate from plasma the platelet surface substrate for thrombin, glycoprotein V (GPV), because a GPV antigen was reported to be present in plasma (3). Plasma fractionation based on procedures for purification of GPV from platelets revealed a thrombin-sensitive protein with appropriate electrophoretic mobility. The protein was purified; an antiserum against it i) reacted with detergent-solubilized platelet proteins or secreted proteins in a double diffusion assay, ii) adsorbed a protein from the supernatant solution of activated platelets, and iii) inhibited thrombin-induced platelet activation, but the antiserum did not adsorb labeled GPV. The purified protein was immunochemically related to prothrombin rather than to GPV. Other antibodies against prothrombin were also able to adsorb a protein from platelets. It is concluded that 1) plasma does not contain appreciable amounts of GPV, and 2) platelets contain prothrombin or an immunochemically similar protein.

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Detection of dengue virus in Aedes mosquitoes in Delhi, India

ENTOMON ◽

10.33307/entomon.v44i3.466 ◽

2019 ◽

Vol 44 (3) ◽

pp. 213-218

Author(s):

Suresh Chand Kaushik ◽

Sukhvir Singh ◽

Purnima Srivastava ◽

R. Rajendran

Keyword(s):

Early Intervention ◽

Early Detection ◽

Aedes Aegypti ◽

Dengue Virus ◽

Infection Rate ◽

Transmission Season ◽

Aedes Mosquitoes ◽

Alternative Approach ◽

Human Sera ◽

Minimum Infection Rate

Detection of viruses in human sera particularly in endemic areas is cumbersome and laborious. Therefore, an alternative approach, Immuno-fluorescence assay (IFA) was performed to determine dengue virus (DENV) positivity in mosquitoes. A total of 1055 adult Aedes aegypti female mosquitoes were tested for IFA test against DENV. Minimum infection rate (MIR) for DENV was found higher during August to November 2016 ranging from 10.75 to 20.83. The average yearly MIR was about 6.64. Higher MIR for Ae. aegypti was found in Sarfabad, Noida (12.71) and Khoda Colony, Ghaziabad (11.90). Minimum MIR (4.67) was observed in Sanjay colony (Faridabad). The main contribution of this study resides in the development of a more suitable monitoring system for early detection of viral circulation and to prioritize early intervention in the non-transmission season.

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IMMUNOLOGICAL REACTIVITY OF INSULIN IN HUMAN SERUM

Acta Endocrinologica ◽

10.1530/acta.0.0530673 ◽

1966 ◽

Vol 53 (4) ◽

pp. 673-680 ◽

Cited By ~ 5

Author(s):

Torsten Deckert ◽

Kai R. Jorgensen

Keyword(s):

Normal Subjects ◽

The Other ◽

Human Pancreas ◽

Porcine Pancreas ◽

Crystalline Insulin ◽

Endogenous Insulin ◽

Significant Difference ◽

Human Sera ◽

Normal Human ◽

The One

ABSTRACT The purpose of this study was to investigate whether a difference could be demonstrated between crystalline insulin extracted from normal human pancreas, and crystalline insulin extracted from bovine and porcine pancreas. Using Hales & Randle's (1963) immunoassay no immunological differences could be demonstrated between human and pig insulin. On the other hand, a significant difference was found, between pig and ox insulin. An attempt was also made to determine whether an immunological difference could be demonstrated between crystalline pig insulin and crystalline human insulin from non diabetic subjects on the one hand and endogenous, circulating insulin from normal subjects, obese subjects and diabetic subjects on the other. No such difference was found. From these experiments it is concluded that endogenous insulin in normal, obese and diabetic human sera is immunologically identical with human, crystalline insulin from non diabetic subjects and crystalline pig insulin.

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STUDIES ON SULFATION FACTOR (SF) ACTIVITY OF HUMAN SERUM

Acta Endocrinologica ◽

10.1530/acta.0.xxxv0381 ◽

1960 ◽

Vol XXXV (III) ◽

pp. 381-396 ◽

Cited By ~ 14

Author(s):

Sven Almqvist

Keyword(s):

Human Serum ◽

Normal Serum ◽

Treatment Level ◽

Response Curves ◽

Normal Children ◽

Significant Difference ◽

Human Sera ◽

Normal Humans ◽

Pre Treatment ◽

Dose Levels

ABSTRACT The sulfation factor (SF) activity of human sera has been estimated using a modification of the method of Daughaday et al. (1959). Each assay was statistically evaluated. The method had a mean precision of 0.14 and, used as an assay of GH of human serum, a sensitivity in three pituitary dwarfs of 0.1 to 0.6 μg of HGH/ml of serum. SF activity was found at all ages between 1 month and 75 years. There was a significantly lower mean SF activity below the age of half a year. Three cases of pituitary dwarfism had significantly low SF activities of sera. There was no significant difference between the SF activities of sera from untreated pituitary dwarfs and the sera from normal children below half a year of age. Dose-response curves with large volumes of sera from pituitary dwarfs and small volumes of sera from normal humans had the same slopes. Four mg of HGH prepared according to the method of Li & Papkoff (1956) resulted in a normal serum SF activity in each of the three dwarfs. A significant (P < 0.01) linear relationship was found between the concentration of SF activity of sera from these subjects and the logarithm of the dose of HGH given with dose levels of 1, 2 and 4 mg daily for three days. The decline of serum SF activity to the pre-treatment level following HGH in one dwarf suggested a half life not different from that indicated by others for growth hormone.

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