US 12,742,778 B2Granted

Test to predict and evaluate innate immune responses to infections and methods of treatment thereof

OwnerSamuel B. Reichberginventor

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Abstract

Disclosed herein are methods of determining innate immune response to infections of a subject comprising mixing a subject blood sample comprising plasma with an anticoagulant solution; adding a sample of innate immune cells incubated with one or more stimulants to the mixture to form a cell sample aliquot; incubating the cell sample aliquots with and without the stimulants for at least one hour at an optimal temperature; separating the plasma from the innate immune cells in the aliquot; measuring one or more cytokines in the plasma; calculating a net stimulation of cytokines the cell sample aliquot; and comparing the net stimulation to a reference group of subjects having adequate innate immunity reactivity. Also disclosed are methods of treating a subject having an innate immune response to an infection.

Claims17

Dependent claims

A method of determining innate immune response to a viral infection of a subject comprising:

(a) mixing a subject blood sample comprising plasma with an anticoagulant solution;
(b) separating the mixture of step (a) into two or more sample mixtures;
(c) adding a sample of innate immune cells incubated with one or more lectins selected from the group consisting of phytohemagglutinin, concanavalin A, lentil lectin, snowdrop lectin, ricin, peanut agglutinin, jacalin, hairy vetch lectin, wheat germ agglutinin, elderberry lectin, Maakia amurensis leukoagglutinin, Maakia amurensis hemagglutinin, Ulex europaeus agglutinin, Aleuria aurantia lectin, and a combination thereof to a first sample mixture of step (b) to form a cell sample aliquot;
(d) adding a sample of innate immune cells not incubated with one or more lectins selected from the group consisting of phytohemagglutinin, concanavalin A, lentil lectin, snowdrop lectin, ricin, peanut agglutinin, jacalin, hairy vetch lectin, wheat germ agglutinin, elderberry lectin, Maakia amurensis leukoagglutinin, Maakia amurensis hemagglutinin, Ulex europaeus agglutinin, Aleuria aurantia lectin, and a combination thereof to a second sample mixture of step (b) to form a control sample aliquot;
(e) incubating the cell sample aliquot and the control sample for at least one hour at a temperature in a range of about 36.1° C. to about 37.2° C.;
(f) separating the plasma from the innate immune cells in the cell sample aliquot to form a plasma cell sample;
(g) separating the plasma from the innate immune cells in the control sample aliquot to form a plasma control sample;
(h) measuring one or more cytokines in the plasma cell sample and one or more cytokines in the plasma control sample;
(i) calculating a net stimulation of cytokines in the cell sample aliquot by subtracting the concentration of cytokines in the control sample aliquot from the concentration of cytokines in the cell sample aliquot;
and
(j) comparing the net stimulation to a reference group of subjects having adequate innate immunity reactivity to the viral infection that limits viral invasion in early infection of the viral infection to determine the innate immune response to a viral infection of the subject.

8 more elements

A method of treating a subject having an innate immune response to a viral infection comprising:

(a) mixing a subject blood sample comprising plasma with an anticoagulant solution;
(b) separating the mixture of step (a) into two or more sample mixtures;
(c) adding a sample of innate immune cells incubated with one or more lectins selected from the group consisting of phytohemagglutinin, concanavalin A, lentil lectin, snowdrop lectin, ricin, peanut agglutinin, jacalin, hairy vetch lectin, wheat germ agglutinin, elderberry lectin, Maakia amurensis leukoagglutinin, Maakia amurensis hemagglutinin, Ulex europaeus agglutinin, Aleuria aurantia lectin, and a combination thereof to a first sample mixture of step (b) to form a cell sample aliquot;
(d) adding a sample of innate immune cells not incubated with one or more lectins selected from the group consisting of phytohemagglutinin, concanavalin A, lentil lectin, snowdrop lectin, ricin, peanut agglutinin, jacalin, hairy vetch lectin, wheat germ agglutinin, elderberry lectin, Maakia amurensis leukoagglutinin, Maakia amurensis hemagglutinin, Ulex europaeus agglutinin, Aleuria aurantia lectin, and a combination thereof to a second sample mixture of step (b) to form a control sample aliquot;
(e) incubating the cell sample aliquot and the control sample for at least one hour at a temperature in a range of about 36.1° C. to about 37.2° C.;
(f) separating the plasma from the innate immune cells in the cell sample aliquot to form a plasma cell sample;
(g) separating the plasma from the innate immune cells in the control sample aliquot to form a plasma control sample;
(h) measuring one or more cytokines in the plasma cell sample and one or more cytokines in the plasma control sample;
(i) calculating a net stimulation of cytokines in the cell sample aliquot by subtracting the concentration of cytokines in the control sample aliquot from the concentration of cytokines in the cell sample aliquot;
(i) comparing the net stimulation to a reference group of subjects having adequate innate immunity reactivity to the viral infection that limits viral invasion in early infection of the viral infection;
and
(k) administering a therapeutic to the subject identified as having the innate immune response to the viral infection.

9 more elements

Dependencies are inferred from the claim text.

Facts

Application: 17/980,235Filed 3 Nov 2022 · us provisional application
Priority: 3 Nov 2021
Published: US 2023/0135111 A14 May 2023
Granted: 22 Sep 2026
Term ends: Not estimatedThe source records don’t support an estimate.

Classification9 codes

G01N 33/569
Immunoassay; Biospecific binding assay; Materials therefor: for microorganisms, e.g. protozoa, bacteria, viruses
A61K 38/21
Cytokines; Lymphokines; Interferons: interferons [IFN]
G01N 33/5047
Involving specific cell types: cells of the immune system
G01N 33/6863
Involving proteins, peptides or amino acids: cytokines, i.e. immune system proteins modifying a biological response such as cell growth…