JP2019070571A - Indication of reactivity of antigen expression and c-myc determination method of gene translocation - Google Patents

Abstract

To provide an indication of reactivity of antigen expression facilitating absolute negative/high probability positive detection of c-MYC gene translocation, and a c-MYC determination method of gene translocation.SOLUTION: Reactivities of antigen expression by immunohistochemistry of c-myc protein are classified into at least 3 types: positive, weak positive and negative. The area ratio of each type of reactivities in the observation area with an optical microscope is calculated. Reactivity symbols indicate each type of the reactivities. Ratio symbol indicates the area ratio of each type of the reactivities to the other types of reactivity. Display symbol displayed while associating the reactive symbol with the ratio symbol is indicated to a displayed item, and an indication of reactivity of antigen expression is created. When the respective items occupy approximately the same one third; i.e., the observation area of positive is 30% to 35%; the observation area of weak positive is 30% to 35%; and the observation area of negative is 30% to 35%, the c-MYC gene translocation is determined that the locus is negative.SELECTED DRAWING: Figure 1

Description

  The present invention relates to indicia of reactivity of antigen expression for immunohistochemistry results and methods of determining c-MYC gene translocation.

Conventionally, pathologists individually have two elements, ie, immunoreactivity (positive, weak positive, negative, etc.) for the result of immunohistochemistry, and a rough proportion (proportion) occupied by them. There is no common notation for showing both together. For this reason, unlike other medical data which often show the result as an absolute value, numerical data was not compared with each other.
For example, as an in vitro method for identifying malignant cells from hematologic malignancies in a sample selected from the group consisting of bone marrow, blood and lymph nodes, type 1 5-HTR and / or type 2 5- A method of detecting HTR-expressing cells by immunocytochemistry is disclosed (see, for example, Patent Document 1).

Japanese Patent Application Publication No. 2017-526629

  Also in the method of Patent Document 1, the method of describing the result of immunohistochemistry is not shown, and a pathologist individually divides the reactivity of the antigen expression into two types, positive and negative, and sensitarily the area ratio Since capturing and identifying malignant cells in a language, there is a problem that the accuracy and reproducibility of identification and the image assumption are bad.

  The present invention was made in view of such conventional problems, and it is an expression of reactivity of antigen expression that facilitates negative / high probability positive determination of translocation of c-MYC gene and c- An object of the present invention is to provide a method of determining MYC gene translocation.

  In order to achieve the above object, the expression of reactivity of antigen expression according to the present invention comprises a reactivity symbol indicating a predetermined reactivity of antigen expression by immunohistochemistry and other reactivity of the predetermined reactivity. It is characterized by having a display symbol displayed in association with a ratio symbol indicating an area ratio to.

  The expression of the reactivity of the expression of the antigen according to the present invention, when the reactivity of the expression of the antigen by immunohistochemistry of the c-myc protein is classified into at least three types of positive, weak positive and negative. It is preferable to have a display symbol which is displayed in association with a symbol of reactivity and a ratio symbol indicating an area ratio of each type of reactivity to other reactivity.

  The classification of reactivity of antigen expression may be four or five or more in addition to three. In the case of four types, for example, “+” for positive, “(weak) +” for weak positivity, and “(very weak) +” for very weak positivity as reactivity symbols indicating predetermined reactivity of antigen expression. And negative signs with "-". "Positive" usually means strong positive.

  The ratio symbol indicating the area ratio is, for example, “>” when the reactivity represented by the reactivity symbol on the left is 60% to 70%, and the reactivity represented by the reactivity symbol on the left is 75% to 90% When ">>" and the reactivity shown by the reactivity symbol is 99% or more on the left side, the symbol shown by ">>>" is mentioned.

In the present invention, the indication may be anything that can display a symbol, and examples include printed matter and other paper, blackboard, monitor image, slide projection image, and the like.
The negative / positive determination of the translocation of c-MYC gene can be facilitated by using the display of the reactivity of the antigen expression according to the present invention.

  The method of determining c-MYC gene translocation according to the present invention classifies the reactivity of antigen expression by immunohistochemistry of c-myc protein into at least three types of positive, weak positive and negative, and occupies each of the observation areas. A display symbol in which area ratios of types of reactivity are determined, and a reactivity symbol indicating each type of reactivity is associated with a ratio symbol indicating an area ratio of each type of reactivity to other types of reactivity The above-mentioned display of reactivity of antigen expression is made by displaying on the subject, the positive is 30 to 35% of the observation area, the weak positive is 30 to 35% of the observation area, the negative is It is characterized in that when it occupies 30 to 35% of the observation area, it is determined that the translocation of the c-MYC gene is negative.

  Another determination method of c-MYC gene translocation according to the present invention classifies the reactivity of antigen expression by immunohistochemistry of c-myc protein into at least three types of positive, weak positive and negative, The area ratio of each type of reactivity is determined, and the reactivity symbol indicating each type of reactivity is displayed in association with the ratio symbol indicating the area ratio of each type of reactivity to the other type of reactivity. The indicator is displayed on the subject to make the aforementioned indicator of the reactivity of the antigen expression, and when the positive is 75 to 90% of the observation area, the translocation of the c-MYC gene becomes positive. It is characterized in that it is determined that the probability of obtaining is high.

In the method of determining c-MYC gene translocation according to the present invention, the present inventors classify the reactivity of the antigen expression by immunohistochemistry of c-myc protein into three types of positive, weak positive and negative, and clinical data It is a judgment method obtained empirically by analyzing
According to the method of determining c-MYC gene translocation according to the present invention, when a predetermined condition is satisfied, negative / positive determination of translocation of c-MYC gene can be performed with high accuracy without performing a precise test. It becomes.
In the present invention, the “observation region” may be the entire photographed micrograph or a partial region specifying a range.

  According to the present invention, it is possible to provide a representation of the reactivity of antigen expression and a method for determining c-MYC gene translocation that facilitates negative and high probability positive determination of translocation of c-MYC gene. .

It is a flowchart which shows the notation method of the representation of the reactivity of the antigen expression of embodiment of this invention. 4 is four photomicrographs of immunohistochemically stained lesion biopsies.

Hereinafter, an embodiment of the present invention will be described based on the drawings.
FIG. 1 is a flow chart showing a method of displaying a representation of reactivity of antigen expression according to the embodiment of this invention. FIG. 2 shows four photomicrographs of immunohistochemically stained lesion biopsies.

  In order to identify various proteins in malignant cells derived from malignant tumors and normal or pathological cells, it is necessary to determine whether the reactivity of antigen expression is positive or negative from photomicrographs of immunohistochemically stained lesion biopsies. It has been done. Preparation of photomicrographs of lesion biopsies and evaluation of reactivity of antigen expression are performed as follows.

  First, the lesion site is collected, and after formalin fixation, paraffin sections are prepared as a mold. Next, paraffin sections are immunohistochemically stained using an anti-c-myc antibody as a primary antibody. The stained sample is observed with a microscope, and the reactivity of the antigen expression by immunohistochemistry of c-myc protein is indicated by a display symbol, and observed with an optical microscope of an eyepiece 10 × objective 40 × lens.

  The reactivity is evaluated from the display symbol of the reactivity of antigen expression, and the degree of expression of c-myc protein in malignant cells is positive, weak positive, very weak positive (weak positive) as the translocation of c-MYC gene The intensity between negatives) and negatives are observed separately, and the area ratio of each antigen expression is symbolized according to the flow chart shown in FIG. An object to be marked may be a monitor image of a personal computer in addition to a predetermined sheet.

  As shown in FIG. 1, when expressing a representation of the reactivity of antigen expression, the area occupied in the observation area is divided into two by “Are well over 50% of the reactivity pattern?”, And the case of yes , "One or two types or three types of reactivity patterns", divided into three, and one type, "+", "(weak) +", "(very weak) +", "-" Mark one of the reactivity symbols as the display symbol.

  The reactivity symbols “+”, “(weak) +”, “(very weak) +” and “−” indicate that the reactivity of the antigen expression by immunohistochemistry of c-myc protein is classified into four types. Each type of reactivity is shown. Of the predetermined reactivity of antigen expression by immunohistochemistry, “+” is positive, “(weak) +” is weak, “(very weak) +” is very weak, “-” is It shows negative. In the photomicrograph of the immunohistochemically stained color of the lesion biopsy, positive is brown, weak positive is light brown, very weak positive is fairly light brown (beige), negative is completely white It is reflected.

  When there are two types of reactivity patterns, “>” and “>>” are between two types of reactivity symbols among “+”, “(weak) +”, “(very weak) +”, and “−”. "," ">>>", ">>> *" display the ratio symbol. As shown in FIG. 1, depending on the area ratio of the reactivity of the two types of reactivity symbols, the left reactivity reactivity symbol is displayed on the left side and the lower reactivity reactivity symbol is displayed on the right side, the ratio Let the symbol be a symbol that is associated with two reactive symbols.

  When there are three types of reactivity patterns, “>”, “(weak) +”, “(very weak) +”, “−” among the reactivity symbols of the three reactivity symbols “>” "," Display the ratio symbol of ">>". As shown in FIG. 1, depending on the area ratio of reactivity of the three types of reactivity symbols, the left reactivity reactivity symbol is displayed on the left side, and the lower reactivity reactivity symbol is displayed on the right side, the ratio Let the symbol be a symbol that is associated with the three reactive symbols.

  In the case of no, if "no about 50% of the reactivity pattern is well over 50%", it is divided into two according to "is about 50% of the reactivity pattern", and if yes, "+", "(weak The ratio of "/", ">", ">>", ">>>" between two or three types of reactivity symbols among +), "(very weak) +" and "-" Display the symbol, and make the ratio symbol a display symbol associated with two or three types of reactive symbols. The ratio symbol "/" indicates that the area ratio of the reactivity of the stronger reactivity symbol on the left to the reactivity of the reactivity symbol of the weaker reactivity on the right is 47-53%.

  When "no", "is there about 50% of the reactivity pattern", "+", "(weak) +", "(very weak) +", and "-" The ratio symbols of “/”, “>>”, “>>>” are displayed in between, and the ratio symbols are displayed symbols associated with the three types of reactivity symbols. The ratio symbol “/” displayed between the three types of reactivity symbols indicates that the area ratio of reactivity of each reactivity symbol is 30 to 35%.

  In the photomicrographs shown in FIG. 2, when the reactivity of antigen expression in each photograph is indicated by a symbol, A is “+ >> (weak) +” and B is “+ / (weak) + >> −” , C is "+ / (weak) +/-", and D is shown by "-".

  Lesion sites were collected, paraffin sections were prepared after formalin fixation, and paraffin sections were immunohistochemically stained using anti-c-myc antibody as a primary antibody. The stained sample was observed with a microscope, and the reactivity of the antigen expression by immunohistochemistry of c-myc protein was indicated by a display symbol according to the flowchart shown in FIG. In addition, using the probe set for FISH (Fluorescence in situ hybridization), positive (including split or fusion with other genes) and negative (without split or fusion with other genes) translocation of c-MYC gene Detected.

  The results are as follows. On the left side, display symbols are shown according to the flowchart. The number of c-MYC gene translocation positive cases in the number of searched cases detected using the probe set for FISH is shown on the right side of each description. Also, on the right side of groups A, B and C, the percentages are shown in parentheses together with the number of similar cases.

Group A: 0/43 (0%)
c-myc + / (weak) +/- · · · 0/43
Group B: 13/25 (52%)
c-myc + / (weak) + >>-... 3/4
c-myc + / (weak) + >>>-... 1/2
c-myc +> (weak) +>-... 0/1
c-myc +> (weak) + >>-... 3/9
c-myc + >> (weak) +>-... 6/9
Group C: 8/9 (89%)
c-myc + >> (weak) + ... 7/7
c-myc + >>> (weak) + ... 1/2

  Thus, when the result of immunohistochemistry is indicated by the symbol "c-myc + / (weak) +/-", positive is 30 to 35% of the observation area and weak positive is 30 to 35% of the observation area Negative, 30 to 35% of the observation area, respectively, occupy approximately the same third. In this case, among 43 cases, 0 cases were positive for translocation of c-MYC gene, and all 43 cases were negative. From this, when the result of immunohistochemistry is "c-myc + / (weak) +/-", it can be determined that the translocation of the c-MYC gene is negative.

  Moreover, when the result of immunohistochemistry is shown by the display symbol "c-myc + >> (weak) +", positive occupies 75 to 90% of the observation area. In this case, among the seven cases, all seven cases were positive for translocation of the c-MYC gene and none were negative. From this, when the result of immunohistochemistry is “c-myc + >> (weak) +”, it can be determined that the translocation of c-MYC gene can be positive with a high probability although the number of cases is small. it can.

  When indicated by the symbol "c-myc + >> (weak) +", that is, when the positive occupies 75 to 90% of the observation area, the probability that the gene A is abnormal is 90% or more. By showing the expression mode at the protein level with the above-mentioned symbols, detection of genetic abnormalities is facilitated. Since it is already known that a patient's prognosis will be poor if there is an abnormality in gene A, it is possible to estimate the patient's prognosis in a protein expression mode by immunohistochemistry.

  Thus, when the result of immunohistochemistry is indicated by the symbol "c-myc + / (weak) +/-", it can be judged negative for translocation of c-MYC gene without detailed examination. Become. In addition, when indicated by the symbol “c-myc + >> (weak) +”, the probability of positive determination of translocation of c-MYC gene is considerably high by using a representation of reactivity of antigen expression Can be done easily.

The above symbols make the immunohistochemistry findings, which so far have been written by different pathologists at different lengths, so as to be considerably shorter according to a certain rule, which makes them easier to read and to compare with each other. It will also shorten the description time. In addition, conventionally, the reaction of antigen expression in two steps of positive and negative is at least in three steps of positive and weak positive and negative etc. c-MYC gene translocation negative / positive The accuracy of the determination can be improved.
Furthermore, handling the findings of immunohistochemistry as big data enables analysis and analysis of large-scale cases.

Claims (4)

  It is characterized by having a display symbol in which a reactivity symbol indicating a predetermined reactivity of antigen expression by immunohistochemistry and a ratio symbol indicating an area ratio to the other reactivity of the predetermined reactivity are associated with each other. Indication of the reactivity of antigen expression.   When the reactivity of the antigen expression by immunohistochemistry of c-myc protein is classified into at least positive, weak positive and negative at least three types, the reactivity symbol showing each type of reactivity and the other types of reactivity A symbol of reactivity of antigen expression characterized by having a display symbol displayed in association with a ratio symbol indicating an area ratio to reactivity.   The reactivity of the antigen expression by immunohistochemistry of c-myc protein is classified into at least three types of positive, weak positive and negative, the area ratio of each type of reactivity occupying the observation area is determined, and each type of reactivity The antigen expression according to claim 2, wherein a display symbol displayed in association with a symbol indicating the reactivity and a ratio symbol indicating the area ratio of each type of reactivity to the reactivity of the other type is displayed on the subject When the positive represents 30 to 35% of the observation area, the weak positive 30 to 35% of the observation area, and the negative 30 to 35% of the observation area; A method for determining c-MYC gene translocation, which comprises determining that the translocation of c-MYC gene is negative. The reactivity of the antigen expression by immunohistochemistry of c-myc protein is classified into at least three types of positive, weak positive and negative, the area ratio of each type of reactivity occupying the observation area is determined, and each type of reactivity The antigen expression according to claim 2, wherein a display symbol displayed in association with a symbol indicating the reactivity and a ratio symbol indicating the area ratio of each type of reactivity to the reactivity of the other type is displayed on the subject And producing a sign of reactivity, and determining that the probability that the translocation of the c-MYC gene can be positive is high when the positivity is 75 to 90% of the observation area. Determination method of MYC gene translocation.