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Hyperthermia augmented in-vitro immune recognition

A hyperthermia and immune response technology, applied in the field of hyperthermia-enhanced in vitro immune recognition, can solve the problem of decreased sensitivity and achieve the effects of increased sensitivity, improved sensitivity, and improved testing and diagnosis

Inactive Publication Date: 2013-03-20
ВИДОУРЕ ХОСПИТАЛ
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0020] Unfortunately, the sensitivity of these tests is reduced in immunocompromised individuals (eg, HIV-infected individuals or patients taking immunosuppressive drugs)

Method used

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  • Hyperthermia augmented in-vitro immune recognition
  • Hyperthermia augmented in-vitro immune recognition
  • Hyperthermia augmented in-vitro immune recognition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0355] Embodiment 1 (preliminary research)

[0356] Elevated temperature enhanced IP-10 and IFN-γ responses.

[0357] We investigated whether IP-10 and IFN-γ responses could be enhanced by increasing the incubation temperature.

[0358] result

[0359] figure 1 is a spaghetti diagram showing two representative individuals from a preliminary study, and we were able to replicate the findings of these experiments in 11 other individuals. We found a consistent and significant increase in signal up to a temperature of 39.5°C, whereas at higher temperatures we observed a sharp drop in response.

[0360] In conclusion, incubation under hyperthermic conditions resulted in increased IP-10 and IFN-γ responses when whole blood from BCG-immunized individuals was stimulated with BCG-specific antigens compared to incubation at 37°C.

[0361] Incubation at high temperature has potential as an easy way to reduce the number of false negative and inconclusive test results and thus improve ...

Embodiment 2

[0362] Embodiment 2 (BCG vaccine research)

[0363] Donors were classified as non-responders and responders by measuring the levels of plasma IP-10 induced by stimulation of whole blood with a M. tuberculosis-specific antigen (TB10.4).

[0364] A total of 35 donors were tested: 16 donors who were BCG immunized and therefore likely to respond to the TB10.4 antigen and 19 non-vaccinated donors who were less likely to respond to the TB10.4 antigen . Whole blood was drawn and incubated for 18 hours at 37°C and 39°C in the presence of the combination of peptides from the antigen TB10.4, respectively.

[0365] Based on preliminary adjustments, we chose to compare the standard 37°C incubation with the new 39°C incubation. We chose to choose the significantly poorer range of 39°C to 39.5°C because we needed proof of concept data for a stable system.

[0366] result

[0367] We collected whole blood from 34 healthy donors (one BCG-immunized donor was excluded due to failed venipunc...

Embodiment 3

[0373] Hyperthermic incubation in the presence or absence of IL-7 and / or anti-IL-10 increased the IP-10 response.

[0374] We verified the effect of hyperthermic incubation on IP-10 responses by testing samples taken from the above 34 responders and non-responders.

[0375] Next, we tested whether the presence of the cytokine IL-7 during incubation enhanced the response. We also tested whether antagonism of the inhibitory cytokine IL-10 would affect the IP-10 response by inhibiting a known major inhibitor in the system (ie inhibitor of inhibition).

[0376] result

[0377] Figure 3A-Figure 3D In column 8 the IP-10 replies are shown. The first 4 columns indicate incubation at 37°C, the last 4 indicate incubation at 39°C. By dividing blood from study participants into equal aliquots and subjecting them to all of the various incubation conditions, direct column to column comparisons can be made. Background levels (blank) were subtracted from antigen and mitogen responses. ...

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Abstract

The present invention relates to a method for generation of a test-antigen specific cell-mediated immune response by incubating at hyperthermic conditions and, more particularly, a method for generation of a test-antigen specific cell-mediated immune response by incubating at hyperthermic conditions and optionally adding IL-7 and / or blocking IL-10. Even more particularly, the present invention provides a method for generating a cell-mediated response to an antigen using whole blood or other suitable biological samples. The method is useful in for immune diagnosis of many infectious diseases, as a marker of immunocompetence, and for detection of T-cell responses to non-self antigens (i.e. infections and vaccines).

Description

technical field [0001] The present invention relates to a general method for generating a test antigen-specific cell-mediated immune response by incubation under hyperthermic conditions; and / or a method of blocking IL-10 to generate a test antigen-specific cell-mediated immune response. More specifically, the invention provides a method of using whole blood or other suitable biological samples to generate a cell-mediated response to an antigen. The method can be used in therapeutic and diagnostic protocols for human, domestic and veterinary, and wild animal use. [0002] Measurement of cell-mediated immune responses is important for the immunodiagnostics of many infectious diseases (as a marker of immune activity) and the detection of T cell responses to non-self antigens (ie infections and vaccines). [0003] The present invention provides a method for producing and / or evaluating a test antigen in a mammal by incubating a sample comprising immune system cells from a mammal ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N33/569G01N33/68
CPCG01N2333/35G01N33/5306G01N33/569G01N33/574G01N33/5047G01N33/6863A61P31/04A61P31/12
Inventor 莫滕·茹华德杰斯珀·欧根-奥尔森佩妮莱·拉温马丁·格罗索斯·阿巴
Owner ВИДОУРЕ ХОСПИТАЛ
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