Process for assessing risk of substance administration
AssigneesR.J. Reynolds Tobacco Company, Wake Forest University Health Sciences
InventorsGaddamanugu L. Prasad, Subhashini Arimilli
Life of this patent
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Abstract
A process for assessing risk that administration of a substance will suppress immune competence in a population of subjects is provided. Such a substance may be a substance related to, contained in or derived from a tobacco product or, more generally, a substance concerning which it is desired to assess risk of administration of the substance.
Claims17
A process for assessing risk that administration of a tobacco product preparation (TPP) substance derived from tobacco will suppress immune competence in a population of subjects, the process comprising:
- collecting a sample of blood from each a plurality of subjects, said subjects being members of the population;
- isolating peripheral blood mononuclear cells (PBMCs) from the sample of blood to form isolated PBMCs;
- administering the TPP substance to the isolated PBMCs for 3 hours, thereby forming substance-exposed PBMCs;
- stimulating the substance-exposed PBMCs with a Toll-like receptor ligand to form Toll-like receptor-ligand stimulated substance-exposed PBMCs, wherein the stimulating takes place for a duration of less than 6 hours;
- measuring one or more levels of a cytokine in the Toll-like receptor-ligand stimulated substance-exposed PBMCs to form one or more measured responses;
- aggregating the one or more measured responses from each of the plurality of subjects to form one or more response aggregates;
- comparing the one or more response aggregates to one or more immune-competence-associated reference ranges measured in control cells,
- thereby assessing risk that administration of the substance will suppress immune competence in the population of subjects.
A process for assessing risk that administration of a tobacco product preparation (TPP) substance derived from tobacco will suppress immune competence in a population of subjects, the process comprising:
- collecting a sample of blood from each a plurality of subjects, said subjects being members of the population;
- isolating peripheral blood mononuclear cells (PBMCs) from the sample of blood to form isolated PBMCs;
- administering the TPP substance derived from tobacco to the isolated PBMCs for 3 hours, thereby forming substance-exposed PBMCs;
- stimulating the substance-exposed PBMCs with phorbol 12-myristate 13-acetate (PMA) and Ionomycin (Iono) to form PMA/Iono stimulated substance-exposed PBMCs, wherein the stimulating takes place for a duration of less than 6 hours;
- measuring one or more responses of levels of a cytokine in the PMA/Iono stimulated substance-exposed PBMCs to form one or more measured responses;
- aggregating the one or more measured responses from each of the plurality of subjects to form one or more response aggregates;
- comparing the one or more response aggregates to one or more immune-competence-associated reference ranges measured in control cells,
- thereby assessing risk that administration of the substance will suppress immune competence in the population of subjects.
A process for assessing risk that administration of a tobacco product preparation (TPP) substance derived from tobacco will suppress immune competence in a population of human subjects, the process comprising:
- collecting a sample of blood from each a plurality of human subjects, said subjects being members of the population;
- isolating peripheral blood mononuclear cells (PBMCs) from the sample of blood to form isolated PBMCs;
- administering the TPP substance to the isolated PBMCs for 3 hours, thereby forming substance-exposed PBMCs;
- stimulating the substance-exposed PBMCs with a Toll-like receptor ligand to form Toll-like receptor-ligand stimulated substance-exposed PBMCs, wherein the stimulating takes place for a duration of less than 6 hours;
- measuring one or more levels of a cytokine in the Toll-like receptor-ligand stimulated substance-exposed PBMCs to form one or more measured responses;
- aggregating the one or more measured responses from each of the plurality of subjects to form one or more response aggregates;
- comparing the one or more response aggregates to one or more immune-competence-associated reference ranges measured in control cells,
- thereby assessing risk that administration of the substance will suppress immune competence in the population of human subjects.
A process for assessing risk that administration of a tobacco product preparation (TPP) substance derived from tobacco will suppress immune competence in a population of human subjects, the process comprising:
- collecting a sample of blood from each a plurality of human subjects, said subjects being members of the population;
- isolating peripheral blood mononuclear cells (PBMCs) from the sample of blood to form isolated PBMCs;
- administering the TPP substance to the isolated PBMCs for 3 hours, thereby forming substance-exposed PBMCs;
- stimulating the substance-exposed PBMCs with PMA/Iono stimulated substance-exposed PBMCs, wherein the stimulating takes place for a duration of less than 6 hours;
- measuring one or more levels of a cytokine in the PMA/Iono stimulated substance-exposed PBMCs to form one or more measured responses;
- aggregating the one or more measured responses from each of the plurality of subjects to form one or more response aggregates;
- comparing the one or more response aggregates to one or more immune-competence-associated reference ranges measured in control cells,
- thereby assessing risk that administration of the substance will suppress immune competence in the population of human subjects.
A process for assessing risk that administration of a tobacco product preparation (TPP) substance derived from tobacco will suppress immune competence in a population of subjects, the process comprising:
- collecting a sample of blood from each a plurality of subjects, said subjects being members of the population;
- isolating peripheral blood mononuclear cells (PBMCs) from the sample of blood to form isolated PBMCs;
- cryopreserving the isolated PBMCs at temperature less than zero degrees Celsius;
- warming the isolated PBMCs to a temperature higher than zero degrees Celsius;
- administering the TPP substance to the isolated PBMCs for 3 hours, thereby forming substance-exposed PBMCs;
- stimulating the substance-exposed PBMCs with a Toll-like receptor ligand to form Toll-like receptor-ligand stimulated substance-exposed PBMCs, wherein the stimulating takes place for a duration of less than 6 hours;
- measuring one or more levels of a cytokine in the Toll-like receptor-ligand stimulated substance-exposed PBMCs to form one or more measured responses;
- aggregating the one or more measured responses from each of the plurality of subjects to form one or more response aggregates;
- comparing the one or more response aggregates to one or more immune-competence-associated reference ranges measured in control cells,
- thereby assessing risk that administration of the substance will suppress immune competence in the population of subjects.
A process for assessing risk that administration of a tobacco product preparation (TPP) substance derived from tobacco will suppress immune competence in a population of subjects, the process comprising:
- collecting a sample of blood from each a plurality of subjects, said subjects being members of the population;
- isolating peripheral blood mononuclear cells (PBMCs) from the sample of blood to form isolated PBMCs;
- cryopreserving the isolated PBMCs at temperature −150 degrees Celsius;
- warming the isolated PBMCs to a temperature 37 degrees Celsius;
- administering the TPP substance to the isolated PBMCs for 3 hours, thereby forming substance-exposed PBMCs;
- stimulating the substance-exposed PBMCs with PMA/Iono to form stimulated substance-exposed PBMCs, wherein the stimulating takes place for a duration of less than 6 hours;
- measuring one or more levels of a cytokine in the PMA/Iono stimulated substance-exposed PBMCs to form one or more measured responses;
- aggregating the one or more measured responses from each of the plurality of subjects to form one or more response aggregates;
- comparing the one or more response aggregates to one or more immune-competence-associated reference ranges measured in control cells,
- thereby assessing risk that administration of the substance will suppress immune competence in the population of subjects.
Dependencies are inferred from the claim text.