CN110441507A - A kind of reagent combination, its kit and its method for detecting patients with uterine myoma immune function - Google Patents
A kind of reagent combination, its kit and its method for detecting patients with uterine myoma immune function Download PDFInfo
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Abstract
Reagent combination, its kit and its method that the invention discloses a kind of for detecting patients with uterine myoma immune function, it is related to biomedicine technical field, specifically, in reagent combination, including multiple reagents that the panimmunity functional parameter to following immune factor is detected, immune factor includes: CD3, CD4, CD8, CD45RA, CCR7, CD127, CXCR3, CXCR5, CCR4, CD56, NKG2D, NKP46, γ δ and V δ 2.Reagent combination to the immune function of patients with uterine myoma can effectively accurately detect, and solve the high disadvantage of prior art testing result inaccuracy, testing cost.
Description
Technical field
The present invention relates to biomedicine technical fields, immune for detecting patients with uterine myoma in particular to one kind
Reagent combination, its kit and its method of function.
Background technique
The generation of the immune function of human immunocyte and various diseases, progress have a directly related property, such as tumour
Occur, development process is exactly because of the result that human autoimmune's cell function is lowered or is suppressed for a long time.Women of childbearing age
Fibroid is a kind of benign tumour for betiding reproductive organs, is a kind of common disease in women population.But current antithetical phrase
The generation cause of disease of palace myomata is not also fully aware of, meanwhile, observation of curative effect in clinical treatment, Prognosis scoveillance etc. also lack
Adequately research.
Due to human immune system play the role of in the generation, development of disease it is vital.For example, the hair of tumour
Raw is mainly to kill since the body function cell being abnormal or the cell of mutation cannot be identified by immunocyte, lead to this
The abnormality proliferation of group's mutant cell, to develop into tumour.
Although fibroid belongs to benign tumour, quality of life and fertility generally will not influence, part is with son
The patient of palace myomata can also generate adverse consequences, such as infertile, menorrhalgia, bleeding etc., to influence the life health of women.
This kind of patient usually tracks curative effect and prognosis by the method that ultrasonic imaging or CT are imaged during clinical treatment.
Currently, lacking the method for capableing of quick, accurate and low expense detection patients with uterine myoma prognosis.
Summary of the invention
The reagent combination that the embodiment of the invention provides a kind of for detecting patients with uterine myoma immune function, the reagent set
Closing to the immune function of patients with uterine myoma can effectively accurately detect, and it is inaccurate to solve prior art testing result
Really, the high disadvantage of testing cost.It should be noted that patients with uterine myoma refer to the patient for once suffering from fibroid and/or
The crowd of susceptible fibroid.
Specifically, reagent combination includes the reagent that is detected of multiple pairs of panimmunity functional parameters, is specifically included pair
Multiple reagents that the panimmunity functional parameter of following immune factor is detected；Immune factor include: CD3, CD4, CD8,
CD45RA, CCR7, CD127, CXCR3, CXCR5, CCR4, CD56, NKG2D, NKP46, γ δ and V δ 2.Further, it is immunized
The factor further includes KIR, NKP30 and CD94.
Further, panimmunity functional parameter includes: T cell, T helper cell, killer T cell, double positive T thin
Born of the same parents, initialization CD4+T cell, terminal differentiation CD4+T cell, CD4+T cell is remembered at center, responsiveness remembers CD4+T cell, just
Beginningization CD8+T cell, terminal differentiation CD8+T cell, CD8+T cell is remembered at center, responsiveness remembers CD8+T cell, inactive
Phase specificity terminal differentiation CD8+T cell, Th2, Tfh2, immature NK cell, activated NK, virus infection specific killing
Property NK cell, V δ 2+ positive cell and functionality V δ 2+ positive cell.
Preferably, panimmunity functional parameter further include: the terminal differentiation CD8+T cell of continuous expression virus-specific,
NK cell, the early function of maturation block property NK cell, conventional lethal NK cell；
It is further preferred that the panimmunity functional parameter further include: T cell, Th and Tc ratio, gamma delta T cells, V δ 1
+ positive cell and (V δ 1+/V δ 2+) cell ratio.
The present invention also provides a kind of method for detecting patients with uterine myoma immune function, this method can be with low cost
Ground mode tests and analyzes crowd's (healthy population) of the crowd or susceptible fibroid that infected fibroid, precisely comments
Estimate the immune functional state of examination crowd, so that detected person can take corresponding measure in time, and the detection method is easily grasped
Make, convenient for promoting.
Specifically, this method includes thin by streaming using the reagent combination of above-mentioned patients with uterine myoma immune function assessment
Born of the same parents' instrument analyzes panimmunity functional parameter.Panimmunity functional parameter please refers to the panimmunity function in reagent combination
Index repeats no more.
When the measurement result of panimmunity functional parameter meets measurement standard, sample is determined as healthy population, index
Measurement standard is as follows:
T cell accounts for the 45.76%~77.45% of lymphocyte；Helper T lymphocyte accounts for the 40%~67% of T cell；Killing
Property T cell accounts for the 31%~58% of T cell；Double positive T cells account for the 1%~5% of T cell；Initialization CD4+T cell accounts for CD4T
The 21.0%~51.6% of cell；Terminal differentiation CD4+T cell accounts for the 13.8%~16.8% of cd4 t cell；Remember CD4+ in center
T cell accounts for the 18.2%~49.4% of cd4 t cell；Responsiveness memory CD4+T cell account for cd4 t cell 27.6%~
55.9%；Initialization CD8+T cell accounts for the 35.4%~53.2% of cd8 t cell；Terminal differentiation CD8+T cell accounts for cd8 t cell
21.2%~40.3%；Center memory CD8+T cell accounts for the 4.8%~6% of cd8 t cell；Responsiveness remembers CD8+T cell
Account for the 33.1%~56.3% of cd8 t cell；Inactive phase specificity terminal differentiation CD8+T cell accounts for terminal differentiation CD8+T cell
85.2%~98.8%；Th2 accounts for the 12%~25% of Th cell；Tfh2 accounts for the 15.3%~23.6% of Tfh cell；Prematurity
NK cell account for the 5.6%~27.8% of NK cell；Activated NK accounts for the 25%~53% of NK cell；Virus infection is specifically killed
Wound property NK cell accounts for the 9.93%~80.85% of NK cell；V δ 2+ positive cell accounts for the 46.8%~72% of gamma delta T cells；Vδ2+
Positive cell accounts for the 94.6%~99.8% of 2 gamma delta T cells of V δ.
Further, the measurement standard of index further include: the terminal differentiation CD8+T cell of continuous expression virus-specific accounts for
The 6.4%~20% of terminal differentiation CD8+T cell；Mature NK cell accounts for the 43~82% of NK cell；Early function blocking property
NK cell accounts for the 36.3%~52.7% of NK cell；Conventional lethal NK cell accounts for the 38.5%~64% of NK cell；
It is further preferred that the measurement standard of index further include: Th and Tc ratio is 0.57~2.44；Gamma delta T cells account for T
The 0.5%~8.2% of cell；V δ 1+ positive cell accounts for the 8.2%~24% of gamma delta T cells；The ratio of (V δ 1+/V δ 2+) is
0.12~0.51.
In addition, the present invention also embodiment additionally provide it is a kind of for detecting the examination of the method for patients with uterine myoma immune function
Agent box comprising the monoclonal antibody that multiple pairs of panimmunity functional parameters are detected；The monoclonal antibody includes:
Anti-human CD3 monoclonal antibody, anti-human CD4 monoclonal antibody, anti-human CD8 monoclonal antibody, anti-human CD 45 RA Dan Ke
Grand antibody, anti-human CCR7 monoclonal antibody, anti-human CD127 monoclonal antibody, anti-human CXCR3 monoclonal antibody, anti-human CXCR5 are mono-
Clonal antibody, anti-human CCR4 monoclonal antibody, anti-human CD56 monoclonal antibody, anti-human NKG2D monoclonal antibody, anti-human NKP46
Monoclonal antibody, anti-human γ δ monoclonal antibody, anti-human 2 monoclonal antibody of V δ, anti-human KIR monoclonal antibody, anti-human NKP30 are mono-
Clonal antibody and anti-human CD94 monoclonal antibody.
Anti-human CD3 monoclonal antibody, anti-human CD4 monoclonal antibody, anti-human CD8 monoclonal antibody, anti-human CD 45 RA Dan Ke
Grand antibody, anti-human CCR7 monoclonal antibody, anti-human CD127 monoclonal antibody, anti-human CXCR3 monoclonal antibody, anti-human CXCR5 are mono-
Clonal antibody, anti-human CCR4 monoclonal antibody, anti-human CD56 monoclonal antibody, anti-human NKG2D monoclonal antibody, anti-human NKP46
Monoclonal antibody, anti-human γ δ monoclonal antibody and anti-human 2 monoclonal antibody of V δ.
Preferably, reagent further include: anti-human KIR monoclonal antibody, anti-human NKP30 monoclonal antibody and anti-human CD94 are mono-
Clonal antibody.
The kit passes through 28 functional parameters of 17 kinds of antibody test samples, can comprehensively reflect exempting from for Women of childbearing age
Epidemic disease functional status, the especially patient with fibroid or the crowd of susceptible fibroid, so that easy infection fibroid
The women that women infected fibroid can take suitably measure in time, maintain health state.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is the first testing result of immune function in the embodiment of the present invention 5；
Fig. 2 is the second testing result of immune function in the embodiment of the present invention 5；
Fig. 3 is the third testing result of immune function in the embodiment of the present invention 5；
Fig. 4 is the 4th testing result of immune function in the embodiment of the present invention 5；
Fig. 5 is the 5th testing result of immune function in the embodiment of the present invention 5；
Fig. 6 is the content of tumor markers CD125 in present invention verifying example 1；
Fig. 7 is the number of the content and myomata of the tumor markers CD125 of sample in present invention verifying example 2.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1
The present embodiment provides it is a kind of detect patients with uterine myoma immune function reagent combination comprising have it is following for pair
The reagent that panimmunity functional parameter is detected, every kind of reagent are respectively corresponded including following any monoclonal antibody；Anti-human CD3
Monoclonal antibody, anti-human CD4 monoclonal antibody, anti-human CD8 monoclonal antibody, anti-human CD 45 RA monoclonal antibody, anti-human CCR7
Monoclonal antibody, anti-human CD127 monoclonal antibody, anti-human CXCR3 monoclonal antibody, anti-human CXCR5 monoclonal antibody are anti-human
CCR4 monoclonal antibody, anti-human CD56 monoclonal antibody, anti-human NKG2D monoclonal antibody, anti-human NKP46 monoclonal antibody resist
People's γ δ monoclonal antibody and anti-human 2 monoclonal antibody of V δ.
Embodiment 2
The present embodiment provides it is a kind of detect patients with uterine myoma immune function reagent combination comprising have it is following for pair
The reagent that panimmunity functional parameter is detected, every kind of reagent are respectively corresponded including following any monoclonal antibody；Anti-human CD3
Monoclonal antibody, anti-human CD4 monoclonal antibody, anti-human CD8 monoclonal antibody, anti-human CD 45 RA monoclonal antibody, anti-human CCR7
Monoclonal antibody, anti-human CD127 monoclonal antibody, anti-human CXCR3 monoclonal antibody, anti-human CXCR5 monoclonal antibody are anti-human
CCR4 monoclonal antibody, anti-human CD56 monoclonal antibody, anti-human NKG2D monoclonal antibody, anti-human NKP46 monoclonal antibody resist
People's γ δ monoclonal antibody, anti-human 2 monoclonal antibody of V δ, anti-human KIR monoclonal antibody, anti-human NKP30 monoclonal antibody and anti-
People's CD94 monoclonal antibody.
Embodiment 3
The present embodiment provides a kind of method for detecting patients with uterine myoma immune function, this method includes using implementation
The reagent combination that example 1 provides, the detection and analysis of panimmunity functional parameter are carried out by flow cytometer to sample.
Panimmunity functional parameter includes: T cell, T helper cell, killer T cell, double positive T cells, initialization
CD4+T cell, terminal differentiation CD4+T cell, center memory CD4+T cell, responsiveness memory CD4+T cell, initialization CD8+T
Cell, terminal differentiation CD8+T cell, center memory CD8+T cell, responsiveness memory CD8+T cell, specificity of inactive phase are eventually
End differentiation CD8+T cell, Th2, Tfh2, immature NK cell, activated NK, virus infection specific killing NK cell, V
δ 2+ positive cell and functionality V δ 2+ positive cell.
When the measurement result of immune function meets measurement standard, the sample is determined as healthy population, the index
Measurement standard it is as follows:
T cell accounts for the 45.76%~77.45% of lymphocyte；Helper T lymphocyte accounts for the 40%~67% of T cell；Killing
Property T cell accounts for the 31%~58% of T cell；Double positive T cells account for the 1%~5% of T cell；Initialization CD4+T cell accounts for CD4T
The 21.0%~51.6% of cell；Terminal differentiation CD4+T cell accounts for the 13.8%~16.8% of cd4 t cell；Remember CD4+ in center
T cell accounts for the 18.2%~49.4% of cd4 t cell；Responsiveness memory CD4+T cell account for cd4 t cell 27.6%~
55.9%；Initialization CD8+T cell accounts for the 35.4%~53.2% of cd8 t cell；Terminal differentiation CD8+T cell accounts for cd8 t cell
21.2%~40.3%；Center memory CD8+T cell accounts for the 4.8%~6% of cd8 t cell；Responsiveness remembers CD8+T cell
Account for the 33.1%~56.3% of cd8 t cell；Inactive phase specificity terminal differentiation CD8+T cell accounts for terminal differentiation CD8+T cell
85.2%~98.8%；Th2 accounts for the 12%~25% of Th cell；Tfh2 accounts for the 15.3%~23.6% of Tfh cell；Prematurity
NK cell account for the 5.6%~27.8% of NK cell；Activated NK accounts for the 25%~53% of NK cell；Virus infection is specifically killed
Wound property NK cell accounts for the 9.93%~80.85% of NK cell；V δ 2+ positive cell accounts for the 46.8%~72% of gamma delta T cells；Vδ2+
Positive cell accounts for the 94.6%~99.8% of 2 gamma delta T cells of V δ.
Embodiment 4
The present embodiment provides a kind of method for detecting patients with uterine myoma immune function, this method includes using implementation
The reagent combination that example 2 provides, the detection and analysis of panimmunity functional parameter are carried out by flow cytometer to sample.
Panimmunity functional parameter includes: T cell, T helper cell, killer T cell, double positive T cells, initialization
CD4+T cell, terminal differentiation CD4+T cell, center memory CD4+T cell, responsiveness memory CD4+T cell, initialization CD8+T
Cell, terminal differentiation CD8+T cell, center memory CD8+T cell, responsiveness memory CD8+T cell, specificity of inactive phase are eventually
End differentiation CD8+T cell, Th2, Tfh2, immature NK cell, activated NK, virus infection specific killing NK cell, V
δ 2+ positive cell, functionality V δ 2+ positive cell, the terminal differentiation CD8+T cell of continuous expression virus-specific, mature NK
Cell, early function blocking property NK cell, conventional lethal NK cell.
When the measurement result of immune function meets measurement standard, the sample is determined as healthy population, the index
Measurement standard it is as follows:
T cell accounts for the 45.76%~77.45% of lymphocyte；Helper T lymphocyte accounts for the 40%~67% of T cell；Killing
Property T cell accounts for the 31%~58% of T cell；Double positive T cells account for the 1%~5% of T cell；Initialization CD4+T cell accounts for CD4T
The 21.0%~51.6% of cell；Terminal differentiation CD4+T cell accounts for the 13.8%~16.8% of cd4 t cell；Remember CD4+ in center
T cell accounts for the 18.2%~49.4% of cd4 t cell；Responsiveness memory CD4+T cell account for cd4 t cell 27.6%~
55.9%；Initialization CD8+T cell accounts for the 35.4%~53.2% of cd8 t cell；Terminal differentiation CD8+T cell accounts for cd8 t cell
21.2%~40.3%；Center memory CD8+T cell accounts for the 4.8%~6% of cd8 t cell；Responsiveness remembers CD8+T cell
Account for the 33.1%~56.3% of cd8 t cell；Inactive phase specificity terminal differentiation CD8+T cell accounts for terminal differentiation CD8+T cell
85.2%~98.8%；Th2 accounts for the 12%~25% of Th cell；Tfh2 accounts for the 15.3%~23.6% of Tfh cell；Prematurity
NK cell account for the 5.6%~27.8% of NK cell；Activated NK accounts for the 25%~53% of NK cell；Virus infection is specifically killed
Wound property NK cell accounts for the 9.93%~80.85% of NK cell；V δ 2+ positive cell accounts for the 46.8%~72% of gamma delta T cells；Vδ2+
Positive cell accounts for the 94.6%~99.8% of 2 gamma delta T cells of V δ；The terminal differentiation CD8+T cell of continuous expression virus-specific accounts for
The 6.4%~20% of terminal differentiation CD8+T cell；Mature NK cell accounts for the 43~82% of NK cell；Early function blocking property
NK cell accounts for the 36.3%~52.7% of NK cell；Conventional lethal NK cell accounts for the 38.5%~64% of NK cell.
Embodiment 5
The present embodiment provides a kind of method for detecting patients with uterine myoma immune function, this method includes using implementation
The reagent combination that example 2 provides, the detection and analysis of 28 immune functions are carried out by flow cytometer to sample.
Panimmunity functional parameter includes: T cell, T helper cell, killer T cell, double positive T cells, initialization
CD4+T cell, terminal differentiation CD4+T cell, center memory CD4+T cell, responsiveness memory CD4+T cell, initialization CD8+T
Cell, terminal differentiation CD8+T cell, center memory CD8+T cell, responsiveness memory CD8+T cell, specificity of inactive phase are eventually
End differentiation CD8+T cell, Th2, Tfh2, immature NK cell, activated NK, virus infection specific killing NK cell, V
δ 2+ positive cell, functionality V δ 2+ positive cell, the terminal differentiation CD8+T cell of continuous expression virus-specific, mature NK
Cell, early function blocking property NK cell, conventional lethal NK cell, T cell, Th and Tc ratio, gamma delta T cells, V δ 1+ sun
Property cell and (V δ 1+/V δ 2+) cell ratio.
When the measurement result of immune function meets measurement standard, the sample is determined as healthy population, the index
Measurement standard it is as follows:
T cell accounts for the 45.76%~77.45% of lymphocyte；Helper T lymphocyte accounts for the 40%~67% of T cell；Killing
Property T cell accounts for the 31%~58% of T cell；Double positive T cells account for the 1%~5% of T cell；Initialization CD4+T cell accounts for CD4T
The 21.0%~51.6% of cell；Terminal differentiation CD4+T cell accounts for the 13.8%~16.8% of cd4 t cell；Remember CD4+ in center
T cell accounts for the 18.2%~49.4% of cd4 t cell；Responsiveness memory CD4+T cell account for cd4 t cell 27.6%~
55.9%；Initialization CD8+T cell accounts for the 35.4%~53.2% of cd8 t cell；Terminal differentiation CD8+T cell accounts for cd8 t cell
21.2%~40.3%；Center memory CD8+T cell accounts for the 4.8%~6% of cd8 t cell；Responsiveness remembers CD8+T cell
Account for the 33.1%~56.3% of cd8 t cell；Inactive phase specificity terminal differentiation CD8+T cell accounts for terminal differentiation CD8+T cell
85.2%~98.8%；Th2 accounts for the 12%~25% of Th cell；Tfh2 accounts for the 15.3%~23.6% of Tfh cell；Prematurity
NK cell account for the 5.6%~27.8% of NK cell；Activated NK accounts for the 25%~53% of NK cell；Virus infection is specifically killed
Wound property NK cell accounts for the 9.93%~80.85% of NK cell；V δ 2+ positive cell accounts for the 46.8%~72% of gamma delta T cells；Vδ2+
Positive cell accounts for the 94.6%~99.8% of 2 gamma delta T cells of V δ；The terminal differentiation CD8+T cell of continuous expression virus-specific accounts for
The 6.4%~20% of terminal differentiation CD8+T cell；Mature NK cell accounts for the 43~82% of NK cell；Early function blocking property
NK cell accounts for the 36.3%~52.7% of NK cell；Conventional lethal NK cell accounts for the 38.5%~64% of NK cell；Continue
The terminal differentiation CD8+T cell of expression virus-specific accounts for the 6.4%~20% of terminal differentiation CD8+T cell；Mature NK is thin
Born of the same parents account for the 43~82% of NK cell；Early function blocking property NK cell accounts for the 36.3%~52.7% of NK cell；Conventional killing
Property NK cell accounts for the 38.5%~64% of NK cell；Th and Tc ratio is 0.57~2.44；Gamma delta T cells account for the 0.5% of T cell
~8.2%；V δ 1+ positive cell accounts for the 8.2%~24% of gamma delta T cells；The ratio of (V δ 1+/V δ 2+) is 0.12~0.51.
The specifying information of method specifically please refers to table 1.
The specifying information of 1 method of table
Patients with uterine myoma and healthy population are detected using the method for embodiment 5, testing result please refers to attached drawing 1
~5.Specifically, attached drawing 1 is the first testing result figure of immune function, and specifically, A is infantilism CD4+T cell in Fig. 1
Testing result, A is center memory-type CD4+T cell detection results in Fig. 1, and C is terminal differentiation effect type CD4+T cell in Fig. 1
Testing result, D is regulatory T cells testing result in Fig. 1, and E is Th2 type T cell testing result in Fig. 1, and F is Tfh type in Fig. 1
T cell testing result；
Fig. 2 is the second testing result of immune function in the embodiment of the present invention 5；Wherein, G is that Tfh1 type T is thin in Fig. 2
Born of the same parents' testing result, H is Tfh2 type T cell testing result in Fig. 2, and I is the cell ratio of Tfh1/Tfh2 in Fig. 2；J is in Fig. 2
Tfh71 type T cell testing result, K is center Memory CD8+ T cells testing result in Fig. 2, and L is terminal differentiation effect in Fig. 2
CD127 high expresses CD8+T cell detection results to type simultaneously；
Fig. 3 is the third testing result of immune function in the embodiment of the present invention 5；Wherein, M is terminal differentiation in Fig. 3
Effect type CD127 low expression CD8+T cell detection results simultaneously, N is Tc17 subtype C D8+T cell detection results, Fig. 3 in Fig. 3
Middle O is total NK cell detection results, and P is the NK cell detection results of CD56 low expression in Fig. 3, and Q is CD94 positive KIR in Fig. 3
Negative NK cell detection results, R is the NK cell detection results of the NKP30 positive in Fig. 3；
Fig. 4 is the 4th testing result of immune function in the embodiment of the present invention 5；Wherein, S is NKG20 positive in Fig. 4
NK cell detection results；T is the NK cell detection results of the NKP46 positive in Fig. 4；U is that total gamma delta T cells detect knot in Fig. 4
Fruit；V is the positive gamma delta T cells testing result of V δ 1 in Fig. 4；W is the positive gamma delta T cells testing result of V δ 2 in Fig. 4；X in Fig. 4
For the cell ratio of V δ 1/V δ 2；
Fig. 5 is the 5th testing result of immune function in the embodiment of the present invention 5；Wherein, Y is NKG2D positive in Fig. 5
2 subgroup gamma delta T cells testing result of V δ；Z is the 2 subgroup gamma delta T cells testing result of V δ of the PD1 positive in Fig. 5.
FIG. 1 to FIG. 5 is the results show that there is significant difference in control group and patient group.
As a result judge:
Reference table 1 to resin be the variation range of cell subsets in healthy population, by immune cell function
The correlation analysis of each parameter and disease generation, development and clinical treatment curative effect, according to parameter difference each between patient and normal person
Different obvious degree is given a mark:
Remarks: statistical term is " conspicuousness ", is indicated with No. *.One star * indicates that the difference reliability is more than 95%,
1 point of note；Two star * * indicate that the difference reliability is more than 99%, remember 2 points；Three star * * * indicate that the difference reliability is more than
99.9%, remember 3 points；
The marking for each parameter that will test is summed, and is obtained total score, is indicated with S, according to total score, with reference to following classification mark
Standard evaluates patient:
S=0 points: normal；
S:0~10 point: normal；
S:11~24 point: immune function is abnormal, need to continue conditioning and restore；
S:25~44 point: immune function is abnormal, pays close attention to, need to improve or therapy intervention；
S:45~54 point: immune function numerous imbalances, it is necessary to keep therapy intervention；
S >=55 point: immune function numerous imbalances have further degradating trend, and palpus intensive treatment is replaced therapeutic scheme or adopted
With immune cell therapy to adjust the associated treatments such as immune function.
Verify example 1
Verify the relationship between the immune function and diagnosing tumor in embodiment 5.
Using the method for embodiment 5, detection and analysis by flow cytometer to sample being adjusted property T cell, by it
Correlation analysis is done with the content of the tumor markers CD125 of sample, the content of tumor markers CD125 please refers to attached drawing 6.
By attached drawing 6 it is found that regulatory T cell number significantly rises in patients with uterine myoma, by itself and tumor markers
The content of CD125 does correlation analysis, and it is in significant positive that discovery regulatory T cells, which rise with the content of tumor markers CD125,
Close (statistics P value is equal to 0.004, shows that correlation is extremely significant).Illustrate that the content of regulatory T cells can be used as uterus muscle
One important indicator of tumor diagnosis and tracing study.
Verify example 2
Verify the relationship between the immune function and diagnosing tumor in embodiment 5.
Using the method for embodiment 5, the detection point of the cell ratio of Tfh1/Tfh2 is carried out to sample by flow cytometer
Analysis, does correlation analysis, tumor markers for the content of tumor markers CD125 and the number of myomata of itself and sample
The content of CD125 and the number of myomata please refer to attached drawing 7.
As shown in Figure 7, the ratio of I type and II type auxiliary type folliculus T cell (Tfh) it is significant in patients with uterine myoma on
Rise, it do with the content of tumor markers CD125 and the number of myomata to correlation analysis, discovery cell ratio rise with
(statistics P value is respectively equal to 0.001 and 0.017, shows that correlation is extremely aobvious in significant positive correlation for CD125 and myomata number
It writes).Illustrate that the ratio of I type and II type auxiliary type folliculus T cell (Tfh) is equally fibroid diagnosis and tracing study and comments
One important indicator of its coincident with severity degree of condition of valence.
Occur the cell subsets of dysfunction in patients, by detecting these parameters, can be diagnosed, be controlled with adjuvant clinical
It treats, especially influence of the evaluation medication process to patient immune function.For example, in therapeutic process, if significantly increasing these
Or reduced parametric regression is normal, therapeutic scheme used by illustrating can restore the immune function of patient, have clinical efficacy.Together
When, if the immune function of patient cannot be made to restore normal by detection discovery treatment, it can also prompt doctor should in time more
Change therapeutic scheme.And after immune function restores normal, then remind doctor that should be discontinued in time.
To sum up, the reagent combination that the embodiment of the invention provides a kind of for detecting patients with uterine myoma immune function, should
In reagent combination, including multiple reagents that the panimmunity functional parameter to following immune factor is detected, immune factor packet
It includes: CD3, CD4, CD8, CD45RA, CCR7, CD127, CXCR3, CXCR5, CCR4, CD56, NKG2D, NKP46, γ δ and V δ
2.Reagent combination to the immune function of patients with uterine myoma can effectively accurately detect, and solve prior art detection
As a result the high disadvantage of inaccuracy, testing cost.Reagent combination can carry out effectively essence to the immune function of patients with uterine myoma
It detects quasi-ly, solves the high disadvantage of prior art testing result inaccuracy, testing cost.
The present invention also provides a kind of method for detecting patients with uterine myoma immune function, this method can be with low cost
Ground mode tests and analyzes crowd's (healthy population) of the crowd or susceptible fibroid that infected fibroid, precisely comments
Estimate the immune functional state of examination crowd, so that detected person can take corresponding measure in time, and the detection method is easily grasped
Make, convenient for promoting.
In addition, in addition, the present invention also embodiment additionally provide it is a kind of for detecting the side of patients with uterine myoma immune function
The kit of method, the kit include the monoclonal antibody that multiple pairs of panimmunity functional parameters are detected；Monoclonal antibody
It include anti-human CD3 monoclonal antibody, anti-human CD4 monoclonal antibody, anti-human CD8 monoclonal antibody, anti-human CD 45 RA monoclonal
Antibody, anti-human CCR7 monoclonal antibody, anti-human CD127 monoclonal antibody, anti-human CXCR3 monoclonal antibody, anti-human CXCR5 Dan Ke
Grand antibody, anti-human CCR4 monoclonal antibody, anti-human CD56 monoclonal antibody, anti-human NKG2D monoclonal antibody, anti-human NKP46 are mono-
Clonal antibody, anti-human γ δ monoclonal antibody, anti-human 2 monoclonal antibody of V δ, anti-human KIR monoclonal antibody, anti-human NKP30 Dan Ke
Grand antibody and anti-human CD94 monoclonal antibody.The kit passes through 28 functional parameters of 17 kinds of antibody test samples, comprehensively
Ground reflects the immune functional state of Women of childbearing age, the especially patient with fibroid or the crowd of susceptible fibroid, with
The women for enabling the women of easy infection fibroid to infect fibroid takes suitably measure in time, keeps body
Health status.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any to repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of reagent for detecting patients with uterine myoma immune function combines, which is characterized in that it includes to following immune
Multiple reagents that the panimmunity functional parameter of the factor is detected；The immune factor include: CD3, CD4, CD8, CD45RA,
CCR7, CD127, CXCR3, CXCR5, CCR4, CD56, NKG2D, NKP46, γ δ and V δ 2.
2. the reagent combination according to claim 1 for detecting patients with uterine myoma immune function, which is characterized in that institute
State immune factor further include: KIR, NKP30 and CD94.
3. the reagent combination according to claim 1 or 2 for detecting patients with uterine myoma immune function, feature exists
In the panimmunity functional parameter includes: T cell, T helper cell, killer T cell, double positive T cells, initialization CD4
+ T cell, terminal differentiation CD4+T cell, center memory CD4+T cell, responsiveness memory CD4+T cell, initialization CD8+T are thin
Born of the same parents, terminal differentiation CD8+T cell, center memory CD8+T cell, responsiveness remember CD8+T cell, inactive phase specificity terminal
Break up CD8+T cell, Th2, Tfh2, immature NK cell, activated NK, virus infection specific killing NK cell, V δ 2
+ positive cell and functionality V δ 2+ positive cell.
4. the reagent combination according to claim 1 or 2 for detecting patients with uterine myoma immune function, feature exists
In the panimmunity functional parameter further include: the terminal differentiation CD8+T cell of continuous expression virus-specific, mature NK
Cell, early function blocking property NK cell, conventional lethal NK cell；
Preferably, the panimmunity functional parameter further include: T cell, Th and Tc ratio, gamma delta T cells, V δ 1+ positive cell
And (V δ 1+/V δ 2+) cell ratio.
5. a kind of for detecting the kit of the method for patients with uterine myoma immune function, which is characterized in that it includes multiple right
The monoclonal antibody that panimmunity functional parameter is detected；The monoclonal antibody includes:
Anti-human CD3 monoclonal antibody, anti-human CD4 monoclonal antibody, anti-human CD8 monoclonal antibody, anti-human CD 45 RA monoclonal are anti-
Body, anti-human CCR7 monoclonal antibody, anti-human CD127 monoclonal antibody, anti-human CXCR3 monoclonal antibody, anti-human CXCR5 monoclonal
Antibody, anti-human CCR4 monoclonal antibody, anti-human CD56 monoclonal antibody, anti-human NKG2D monoclonal antibody, anti-human NKP46 Dan Ke
Grand antibody, anti-human γ δ monoclonal antibody and anti-human 2 monoclonal antibody of V δ.
6. according to claim 5 for detecting the kit of the method for patients with uterine myoma immune function, feature exists
In the monoclonal antibody further includes anti-human KIR monoclonal antibody, anti-human NKP30 monoclonal antibody and anti-human CD94 Dan Ke
Grand antibody.
7. a kind of method for detecting patients with uterine myoma immune function, which is characterized in that any using claims 1 to 3
Described in patients with uterine myoma immune function assessment reagent combination by flow cytometer to panimmunity functional parameter into
Row analysis.
8. the method according to claim 7 for detecting patients with uterine myoma immune function, which is characterized in that described more
Kind immune function includes: T cell, T helper cell, killer T cell, double positive T cells, initialization CD4+T cell, end
End differentiation CD4+T cell, center memory CD4+T cell, responsiveness memory CD4+T cell, initialization CD8+T cell, terminal point
Change CD8+T cell, center memory CD8+T cell, responsiveness and remembers CD8+T cell, inactive phase specificity terminal differentiation CD8+T
Cell, Th2, Tfh2, immature NK cell, activated NK, virus infection specific killing NK cell, V δ 2+ positive cell
And functionality V δ 2+ positive cell；
Preferably, the panimmunity functional parameter further include: the terminal differentiation CD8+T cell of continuous expression virus-specific,
NK cell, the early function of maturation block property NK cell, conventional lethal NK cell；
It is further preferred that the panimmunity functional parameter further include: T cell, Th and Tc ratio, gamma delta T cells, V δ 1+ sun
Property cell and (V δ 1+/V δ 2+) cell ratio.
9. the method according to claim 7 for detecting patients with uterine myoma immune function, which is characterized in that described to exempt from
When the measurement result of epidemic disease functional parameter meets measurement standard, sample is determined as healthy population, and the measurement standard of the index is as follows:
T cell accounts for the 45.76%~77.45% of lymphocyte；Helper T lymphocyte accounts for the 40%~67% of T cell；Lethal T
Cell accounts for the 31%~58% of T cell；Double positive T cells account for the 1%~5% of T cell；It is thin that initialization CD4+T cell accounts for CD4 T
The 21.0%~51.6% of born of the same parents；Terminal differentiation CD4+T cell accounts for the 13.8%~16.8% of CD4 T cell；Remember CD4+T in center
Cell accounts for the 18.2%~49.4% of CD4 T cell；Responsiveness memory CD4+T cell account for CD4 T cell 27.6%~
55.9%；Initialization CD8+T cell accounts for the 35.4%~53.2% of cd8 t cell；Terminal differentiation CD8+T cell accounts for CD8 T cell
21.2%~40.3%；Center memory CD8+T cell accounts for the 4.8%~6% of CD8 T cell；Responsiveness remembers CD8+T cell
Account for the 33.1%~56.3% of CD8 T cell；It is thin that inactive phase specificity terminal differentiation CD8+T cell accounts for terminal differentiation CD8+T
The 85.2%~98.8% of born of the same parents；Th2 accounts for the 12%~25% of Th cell；Tfh2 accounts for the 15.3%~23.6% of Tfh cell；Not at
Ripe NK cell accounts for the 5.6%~27.8% of NK cell；Activated NK accounts for the 25%~53% of NK cell；Virus infection is special
Lethal NK cell accounts for the 9.93%~80.85% of NK cell；V δ 2+ positive cell accounts for the 46.8%~72% of gamma delta T cells；Vδ
2+ positive cell accounts for the 94.6%~99.8% of 2 gamma delta T cells of V δ.
10. the method according to claim 7 for detecting patients with uterine myoma immune function, which is characterized in that described
The measurement standard of index further include: the terminal differentiation CD8+T cell of continuous expression virus-specific accounts for terminal differentiation CD8+T cell
6.4%~20%；Mature NK cell accounts for the 43~82% of NK cell；Early function blocking property NK cell accounts for NK cell
36.3%~52.7%；Conventional lethal NK cell accounts for the 38.5%~64% of NK cell；
Preferably, the measurement standard of the index further include: Th and Tc ratio is 0.57~2.44；Gamma delta T cells account for T cell
0.5%~8.2%；V δ 1+ positive cell accounts for the 8.2%~24% of gamma delta T cells；The ratio of (V δ 1+/V δ 2+) be 0.12~
0.51。
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