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The determination of the cytokines confirms the switching-on
of the
cellular immune answer after the introduction of vaccine RESAN
Under the action of RESAN the cells of the immune
system get activated.
Fig. 1 illustrates a classical
scheme showing how the T-helpers function out.
Fig. 1. Development and interaction
of the T- helpers of 1 and 2 types,
production of cytokines
The prevailing production of the T- helpers of type
1 (Th1) shows an activation of the cellular immune answer, where as
the prevailing production of T- helpers of 2 type (Th2) shows an activation
of the humoral immune answer. On a basis of the contents of the cytokines
in vivo (in blood serum) one can make a conclusion whether the cellular
or the humoral immune answer is prevailing. After the administration
of the vaccine RESAN, an authentic rise in the level of TNF-a
(a factor of necrosis of tumour-alpha) and IFN-g
(gamma-interferon) take place, showing the activation of the cellular
immune answer.
Determination of these cytokines in blood serum is
carried out before and in 7-14 days after the administration of the
drug.
In normal, the blood serum contents 0-50 pg/mls (picogram
per ml) of TNF-a and IFN-g.
A simple test proving the immune responses of the drug based
on the
general blood test
The general blood test is taken prior to the injection
of the drug RESAN
, and through 1,7,14 days after the injection.
On the next day of the injection of RESAN, the growth
of the leucocytes (mainly phagocytes) is observed. An increase of
segmented and rod nuclear cells and a decrease of the lymphocytes
are observed in the leucocyte formula.
After 7 days of injection, the decrease in the number
of leucocytes in compared with the prior analysis is observed. A decrease
in the percentage of segmentated cells, rod nuclear cells and increase
of the lymphocytes are observed in the leucocyte formula.
After 14 days of the injection, the number of leucocytes
becomes same as was prior the injection, where as in the leucocyte
formula an increase of lymphocytes is observed.
The rosette-forming test, which shows
the antitumoral responses of the drug |
The blood tests are done as in the case of marking
the immune status.The tests are carried out before and after the 10-12
days of the injection.
It is necessary to grow a tumor cell culture from
the tumoral tissue obtained during biopsy (in a sterile condition).
The tumor cell suspension containing 6·107 cells/1
ml is prepared. From the blood plasma a leucocyte suspension of 2·106
per ml is prepared. 0.1 ml each of leucocyte suspension and tumor
cell suspension are mixed and stirred in a "U"- shaped plate. The
mixer is incubated for 30 minutes in 370 C and centrifuged
for 3 minutes in 1000 rev/min. The undescended liquid part is removed
and 0.1 ml of 0.05% of glutaraldihyde is added, carefully stirred.
0.1 ml of the solution is replaced in an object-
plate, dried out, fixed up with alcohol and is colored with the solution
of Romanovski-Hinza. The number of rosettes is counted up through
a microscope. The increase in the rosette formation of lymphocytes
with tumor cells states the antitumoral immune responses.
The tumor cell lysis test (in a sterile condition)
The culture of tumoral cells from the tumoral tissues
obtained during the biopsy is grown up. The test is carried out before
and 10-12 days after the injection. A suspension of 2.5·103
cells per ml is prepared from the tumor cell culture. 20 ml of this
suspension is replaced in a (falcon No. 3034, USA). Another suspension
containing T-lymphocytes of 5·105 cells per ml is
prepared from the patient's blood plasma after 24 hrs. The liquid
part above the precipitated layer from the microtitre- plate is removed
out. Around 200 tumor cells are left adhesed on the surface of the
plate. About 20 mcl (microlitre) suspension of T- lymphocytes is added
to them, incubated in 1 atmosphere with 10% CO2 for 48
hrs. After washing 2 times, it is fixed up by alcohol and colored
with solution- Hinza. The number of live tumor cells left are counted
up. The result is calculated using the formula- tumor cell lysis (TCL)
in %, it shows the % lysis of tumoral cells in 48 hrs.
TCL = ( NTCC - NTCE)/ NTCC·100%.
Where NTCC- the number of tumor cells in control,
NTCE- number of tumor cells in experiment. If TCL = 30% and above,
it shows that the antitumor immune system has been switched on.
A conlusion is made after comparing the results obtained
before and after the injection.
The monitoring of the immunotherapy based on the level of the
tumor-associated antigens (tumor markers)
For the verification of effectiveness of the administered
immunotherapy, a monitoring of the tumor-associated antigens (tumor
markers) may be used.
- The maintenance of the dynamic growth of the tumor-associated
antigens in the patient's blood shows the ineffectiveness of the
immunotherapy.
- The slow dinamic growth of the tumor-associated antigens shows
a weak immune responses.
- A halt in the growth or a decrease in the level of the tumor-associated
antigens shows an effective antitumor immune responses.
Here are some accepted blood tumor markers for common cancers:
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The malignant tumors
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Tumor Marker Blood Test
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Tumors of the alimentary tract (esophagus, stomach, colon,
rectum, etc.)
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CEA,
CA 19-9,
CA 195,
CA 72-4,
CA 50,
CMU10,
DUPAN-2,
CO17-1A,
GA733,
GGTP.
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Liver cancers
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AFP,
CEA,
CA 19-9.
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Pancreatic cancer
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CA 19-9,
CEA,
GGTP.
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Lung cancers
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NSE,
CK-BB,
MUC-1,
CA 19-9,
DUPAN-2.
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Breast cancers
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CEA,
CA 15-3,
CA 549,
CAM26,
CA M29,
CA27.29,
MUC-1,
TPS,
BCA225,
317G5,
454C11.
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Ovarian cancers
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CA 125,
MUC-1,
TPS.
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Prostate cancers
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PSA
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Cancers of testis
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AFP,
b-hCG.
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Bone cancers
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TP-3
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If you have any questions concerning
the cancer diagnosis, monitorings you may contact
us.
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