Diagnostic method for Alzheimer's disease

• US 6,300,085 B1
• Filed: 07/18/1997
• Issued: 10/09/2001
• Est. Priority Date: 05/03/1993
• Status: Expired due to Fees

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Original Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Inventors: Alkon, Daniel L.
• Primary Examiner(s): DUFFY, PATRICIA ANN
• Application Number:
  US08/809,646
• Time in Patent Office: 1,544 Days
• Field of Search: 435/7.1, 435/7.21, 435/7.24, 435/7.92, 435/7.94, 435/7.95, 436/518, 436/528, 436/530, 436/531, 436/63, 436/811, 436/172
• US Class Current: 435/7.21
• CPC Class Codes: C07K 14/4711 Alzheimer's disease; Amyloid plaque core protein
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: G01N 2333/9121 with an alcohol group as acceptor (2.7.1), e.g. general tyrosine, serine or threonine kinases
• CPC Class Codes: G01N 2800/2821 Alzheimer
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes
• CPC Class Codes: G01N 33/6872 Intracellular protein regulatory factors and their receptors, e.g. including ion channels
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function

Abstract

The present invention provides methods for the diagnosis of Alzheimer'"'"'s disease using human cells. Specifically, one method detects differences between potassium channels in cells from Alzheimer'"'"'s patient and normal donors, and differences in intracellular calcium concentrations between Alzheimer'"'"'s and normal cells in response to chemicals known to increase intracellular calcium levels. Other methods detect differences between the memory associated GTP binding Cp20 protein levels between Alzheimer'"'"'s and normal cells. In addition a diagnostic index for improved assessment between Alzheimer'"'"'s and non Alzheimer'"'"'s cells is provided.

Claims

1. A method for assaying a sample of viable cells from a patient for the presence of Alzheimer'"'"'s disease, said method comprising the steps of:

i) assaying said sample with a bombesin Alzheimer'"'"'s disease diagnostic test, wherein a score of 0 indicates the absence of Alzheimer'"'"'s disease and a score of −

0.5 or −

1.0 indicates the presence of Alzheimer'"'"'s disease;

ii) assaying said sample with a bombesin Alzheimer'"'"'s disease diagnostic test, wherein a score of 0 indicates the absence of Alzheimer'"'"'s disease and a score of −

0.5 or −

1.0 indicates the presence of Alzheimer'"'"'s disease; and

iii) adding the scores obtained from said bradykinin and a bombesin Alzheimer'"'"'s disease diagnostic tests, wherein a negative value indicates the presence of Alzheimer'"'"'s disease and a positive value indicates the absence of Alzheimer'"'"'s disease.

2. The method according to claim 1, wherein the bradykinin Alzheimer'"'"'s disease diagnostic test comprises contacting the sample with bradykinin for a time period not more than 135 seconds, and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration, wherein a score of zero is assigned when the percentage of responding cells is less than 1.5%, a score of −

0.5 is assigned when the percentage of responding cells is greater than or equal to 1.5%, and a score of −

1 is assigned when the percentage of cells responding is greater than or equal to 4%.

3. The method according to claim 1, wherein the bombesin Alzheimer'"'"'s disease diagnostic test comprises contacting the sample with bombesin for a time period not more than 40 seconds, and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration and the magnitude of response, the magnitude of response being represented by an area under a graph of calcium ion concentration as a function time, wherein a score of zero is assigned when the percentage of responding cells is less than 50% or the area <

23000 nM-sec, a score of −

0.5 is assigned when the percentage of responding cells is greater than or equal to 50% and an area greater than or equal to 2300 nM-sec, and a score of −

1 is assigned when the percentage of cells responding is greater than or equal to 50% and an area greater than or equal to 30000 nM-sec.

4. A method for assaying a sample of viable cells from a patient for the presence of Alzheimer'"'"'s disease, said method comprising the steps of:

i) assaying said sample with a bradykinin Alzheimer'"'"'s disease diagnostic test, wherein a score of 0 indicates the absence of Alzheimer'"'"'s disease and a score of −

0.5 or −

1.0 indicates the presence of Alzheimer'"'"'s disease;

ii) assaying said sample with a TEA Alzheimer'"'"'s disease diagnostic test, wherein a score of 0.5 or 1 indicates the absence of Alzheimer'"'"'s disease and a score of 0 indicates the presence of Alzheimer'"'"'s disease; and

iii) adding the scores obtained from said bradykinin and TEA Alzheimer'"'"'s disease diagnostic tests, wherein a negative value indicates the presence of Alzheimer'"'"'s disease and a positive value indicates the absence of Alzheimer'"'"'s disease.

5. The method according to claim 4, wherein the bradykinin Alzheimer'"'"'s disease diagnostic test comprises contacting the sample with bradykinin for a time period not more than 135 seconds, and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration, wherein a score of zero is assigned when the percentage of responding cells is less than 1.5%, a score of −

0.5 is assigned when the percentage of responding cells is greater than or equal to 1.5%, and a score of −

1 is assigned when the percentage of cells responding is greater than or equal to 4%.

6. The method according to claim 4, wherein the TEA Alzheimer'"'"'s disease diagnostic test comprises contacting said sample with TEA for a time period not more than 60 seconds and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration, wherein a score of zero is assigned when the percentage of responding cells is less than 5%, a score of 0.5 is assigned when the percentage of responding cells is greater than or equal to 5%, and a score of 1 when the percentage of cells responding is greater than or equal to 16%.

7. A method for assaying a sample of viable cells from a patient for the presence of Alzheimer'"'"'s disease, said method comprising the steps of:

i) assaying said sample with a bombesin Alzheimer'"'"'s disease diagnostic test, wherein a score of 0 indicates the absence of Alzheimer'"'"'s disease and a score of −

0.5 or −

1.0 indicates the presence of Alzheimer'"'"'s disease;

ii) assaying said sample with a TEA Alzheimer'"'"'s disease diagnostic test, wherein a score of 0.5 or 1 indicates the absence of Alzheimer'"'"'s disease and a score of 0 indicates the presence of Alzheimer'"'"'s disease; and

iii) adding the scores obtained from said bombesin and TEA Alzheimer'"'"'s disease diagnostic tests, wherein a negative value indicates the presence of Alzheimer'"'"'s disease and a positive value indicates the absence of Alzheimer'"'"'s disease.

8. The method according to claim 7, wherein the bombesin Alzheimer'"'"'s disease diagnostic test comprises contacting the sample with bombesin for a time period not more than 40 seconds, and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration and the magnitude of response, the magnitude of response being represented by an area under a graph of calcium ion concentration as a function of time, wherein a score of zero is assigned when the percentage of responding cells is less than 50% or the area <

23000 nM-sec, a score of −

0.5 is assigned when the percentage of responding cells is greater than or equal to 50% and an area greater than or equal to 23000 nM-sec, and a score of −

1 is assigned when the percentage of cells responding is greater than or equal to 50% and an area greater than or equal to 30000 nM-sec.

9. The method according to claim 7, wherein the TEA Alzheimer'"'"'s disease diagnostic test comprises contacting said sample with TEA for a time period not more than 60 seconds and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration, wherein a score of zero is assigned when the percentage of responding cells is less than 5%, a score of 0.5 is assigned when the percentage of responding cells is greater than or equal to 5%, and a score of 1 when the percentage of cells responding is greater than or equal to 16%.

10. A method for assaying a sample of viable cells from a patient for the presence of Alzheimer'"'"'s disease, said method comprising the steps of:

i) assaying said sample with a bradykinin Alzheimer'"'"'s disease diagnostic test, wherein a score of 0 indicates the absence of Alzheimer'"'"'s disease and a score of −

0.5 or −

1.0 indicates the presence of Alzheimer'"'"'s disease;

ii) assaying said sample with a bombesin Alzheimer'"'"'s diagnostic test, wherein a score of 0 indicates the absence of Alzheimer'"'"'s disease and a score of −

0.5 or −

1.0 indicates the presence of Alzheimer'"'"'s disease; and

iii) assaying said sample with a TEA Alzheimer'"'"'s disease diagnostic test, wherein a score of 0.5 or 1 indicates the absence of Alzheimer'"'"'s disease and a score of 0 indicates the presence of Alzhimer'"'"'s disease; and

iv) adding the scores obtained from said bradykinin, bombesin and TEA Alzheimer'"'"'s disease diagnostic tests, wherein a negative value indicates the presence of Alzheimer'"'"'s disease and a positive value indicates the absence of Alzheimer'"'"'s disease.

11. The method according to claim 10, wherein the bradykinin Alzheimer'"'"'s disease diagnostic test comprises contacting the sample with bradykinin for a time period not more than 135 seconds, and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration, wherein a score of zero is assigned when the percentage of responding cells is less than 1.5%, a score of −

0.5 is assigned when the percentage of responding cells is greater than or equal to 1.5%, and a score of −

1 is assigned when the percentage of cells responding is greater than or equal to 4%.

12. The method according to claim 10, wherein the bombesin Alzheimer'"'"'s disease diagnostic test comprises contacting the sample with bombesin for a time period not more than 40 seconds, and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration and the magnitude of response, the magnitude of response being represented by an area under a graph of calcium ion concentration as a function of time, wherein a score of zero is assigned when the percentage of responding cells is less than 50% or the area <

23000 nM-sec, a score of −

0.5 is assigned when the percentage of responding cells is greater than or equal to 50% and an area greater than or equal to 23000 nM-sec, and a score of −

1 is assigned when the percentage of cells responding is greater than or equal to 50% and an area greater than or equal to 30000 nM-sec.

13. The method according to claim 10, wherein the TEA Alzheimer'"'"'s disease diagnostic test comprises contacting said sample with TEA for a time period not more than 60 seconds and measuring the percentage of cells responding with an increase in intracellular calcium ion concentration, wherein a score of zero is assigned when the percentage of responding cells is less than 5%, a score of 0.5 is assigned when the percentage of responding cells is greater than or equal to 5%, and a score of 1 when the percentage of cells responding is greater than or equal to 16%.