EmCell :: The new method of immune substitution therapy with the use of embryonic cells in HIV infection (AIDS)

THE WORLDS LARGEST CLINICAL EXPERIENCE IN EMBRYONIC STEM CELL TRANSPLANTATION FOR
VARIOUS DISEASES AND CONDITIONS.

Endoscopic Treatment of Liver Diseases

Experience of Application of Embryonic Stem Cells in Amyotrophic Lateral Sclerosis

Embryonic Stem Cells in New-Onset Type 2 Diabetes Mellitus

Embryonic Stem Cells in Pernicious Decompensated Type 2 Diabetes Mellitus

Embryonic Stem Cell Transplantation in Multiple Sclerosis

Embryonic Cell Suspensions in Rheumatoid Arthritis



	16 YEARS OF INTERNATIONAL CLINICAL EXPERIENCE IN TRANSPLANTATION OF HUMAN EMBRYONIC/FETAL STEM CELLS. REVIEW.

The new method of immune substitution therapy with the use of embryonic cells in HIV infection (AIDS)

Protected by U.S.Patent No.5,811,089

Greece Patent No1002969

Other patents pending.

Treatment of AIDS

Case reports

Tables

Treatment of AIDS comprises two major points:

- control of HIV virus, and
- reconstitution of the patient's immunity and general condition.

While in the direction of virus control, considerable progress has been made, but no efficient means is available in the second direction, i.e. reconstitution of immunity.

The above U.S. Patent protects a method allowing the immunity of an organism to be restored substantially and quickly, thereby resulting in significant clinical progress in AIDS treatment. In case of HIV infection, this method helps to prevent or significantly delay the development of AIDS manifestations and AIDS-associated complex.

The method is based on the use of Human Embryonic Cells (HFC). According to studies carried out by European, the U.S. and Russian Patent and Trademark Offices, its novelty has been rated A-level.

The use is made of cryopreserved suspensions containing live Human Fetal Cells which are capable of reproducing inside the human organism, thereby providing reconstitution of hematopoiesis, including its immunological component. Using the patented method there is no need to either solve the problem of histocompatibility or to worry about the risk of patient infection.

Duration of the effects provided by each treatment session ranges from several months to between 1 to 2 years. Positive changes of both laboratory indices of the immune system and, which is especially important, of the clinical picture, are observed. Repeated treatment courses permit us to substantially delay the fatal outcome of the disease.

Antiretroviral Therapy and Cell Therapy may be combined together to achieve the maximum effective treatment. In terms of manifestation of the curing effect, this method has no equals among existing methods of treatment.

Application of Human Embryonic Cells according to the patented method causes no immune conflict, and combines the advantages of living cells transplantation with simple and safe use of a regular medicinal preparation.

The main group consisted of 38 patients for which preliminary selection criteria had been developed: impossibility of using antiviral therapy because of intolerance or considerable side effects; more than 50% decrease in CD4+ lymphocytes during preceding 6 months or 100/1 mm3 reduction during the same time; progress of disease from ARC (AIDS-related complex) to AIDS; worsening of capacity for work and quality of living (decrease in Karnofsky index by more than 20% during last 6 months); actual incurability of a patient (CD4+ amount less than 50/mm3 in manifestation of AIDS-indicator diseases, and with Karnofsky index below 30%). Average age of patients was 36 years (within 23 to 54 years); average time since the date of diagnosis of the infection 5 years; average level of CD4+ lymphocytes 183/mm3.

Elevation of levels of CD4+, CD3+ and CD8+ lymphocytes evidences that Embryonic Stem Cells Transplantation in AIDS patients results in two-phase, i.e. after 2 weeks and 2 months, increase in the level of these parameters, being statistically reliable for CD4+ and CD3+

The percentage of elevation vs initial level is shown

Data of 17 patients for whom multiple studies of the immunity have been performed.

Since the range of initial values of CD4+ lymphocytes is wide, we divided patients into 6 groups according to the initial level of CD4+.

Fig. 1 Average initial CD4+ lymphocytes level in Groups with different initial CD4+ level (cells/ml)

Fig. 2 Number of patients in groups with different initial level of CD4+ lymphocytes

Fig. 3 Average CD4+ level after 7 days

Fig. 4 Percentage of CD4+ elevation after 7 days

Fig. 5 Average CD4+ level after 14 days

Fig. 6 Percentage of CD4+ elevation after 14 days

Fig. 7 Average CD4+ level after 30 days

Fig. 8 Percent of CD4+ after 30 days

Fig. 9 Average CD4+ level after 60 days

Fig. 10 Percent of CD4+ elevation after 60 days

Fig.1 demonstrates average values and standard errors for CD4 in each group prior to treatment.. Fig.2 gives the number of patients in each group. Fig.3 shows the data on CD4 levels in relevant groups 1 week after treatment, and Fig.4 demonstrates percentage of these values with respect to the initial level. Fig. 5 to 10 show the same in further observation terms.

Fig.11 CD4+ lymphocytes (cells/ml)

Fig.12 CD4+ lymphocytes (%)

Fig. 13 CD3+ lymphocytes (cells/ml)

Fig. 14 CD3+ lymphocytes (%)

Fig. 15 CD8+ lymphocytes (cells/ml)

Fig. 16 CD8+ lymphocytes (%)

At Figs 11 and 12, changes of CD4 variations is given without division into groups according to initial levels. After 1 month stimulating effects go and relative decrease of CD4 level is observed. By 2 months after treatment a second wave of CD4 level increase is observed, that is associated with the effect of immune substitution. After 1 and 2 weeks, and 2 months the increase of CD4 level is statistically reliable. Data on CD3 and CD8 evidence reliable increase by 2 weeks and 1 month.

Changes of CD4+ in patients with AIDS related complex (AIDS ARC) after Embryonic Stem Cell Transplantation

Statistic indices	units	N	T	p	Min	Max	Mean	St.err.
Before treatment	abs	26			15	631	232,8	35,5
	%	26			1,1	34,6	12,1	1,8
After 7 days	abs	26	0,0	>0,05	17	821	232,1	39,4
	%	26	0,3	>0,05	1,2	38,0	12,3	1,9
After 14 days	abs	26	2,1	0,042	20	1115	324,9	60,4
	%	26	1,9	>0,05	1,3	35,0	13,9	1,9
After 30 days	abs	26	1,3	>0,05	10	956	281,9	47,7
	%	26	0,5	>0,05	1,2	33,0	12,6	1,7
After 60 days	abs	13	2,3	0,037	15	839	351,5	78,2
	%	13	1,0	>0,05	1,1	23,0	14,9	2,6

Changes of CD4+ in AIDS patients after Embryonic Stem Cell Transplantation

Statistical parameters	units	N	T	p	Min	Max	Mean	St.err.
Before treatment	abs	8			6	189	43,1	21,3
	%	8			1,1	20,0	5,5	2,2
After 7 days	abs	8	2,8	0,026	0	288	80,1	32,4
	%	8	1,4	>0,05	0	19,6	7,4	2,5
After 14 days	abs	8	2,5	0,043	3	272	86,6	32,7
	%	8	0,6	>0,05	0,7	16,6	6,5	1,9
After 30 days	abs	8	0,7	>0,05	3	174	64,6	20,3
	%	8	0,4	>0,05	0,7	16,6	6,4	1,8
After 60 days	abs	3	1,3	>0,05	22	47	36,3	7,4
	%	3	1,3	>0,05	1,6	16,6	7,4	4,7

Changes of CD4+ in AIDS patients with different initial level of Karnofsky index after Embryonic Stem Cell Transplantation

Karnofsky	Stat.	Period of observation after Embryonic Stem Cell Transplantation
Index	index	Before	7 days after	14 days after	30 days after	60 days after
90-100	M	220,4	236,8	295,3	244,5	300,6
	m	44,2	53,2	74,8	60,9	90,5
	p		>0,05	<0,05	>0,05	<0,05
	N	17	17	17	17	11
70-80	M	221,8	220,7	303,8	265,3	255,4
	m	84,0	81,1	175,4	124,4	103,1
	p		>0,05	<0,05	<0,05	>0,05
	N	6	6	6	6	5
40-50	M	150,7	120,1	174,1	208,1	158,5
	m	70,4	49,8	59,7	58,3	136,5
	p		>0,05	<0,05	<0,05	>0,05
	N	7	7	7	7	7
10-20	M	66,5	119,0	133,8	68,0
	m	41,2	59,2	55,9	20,4
	p	>0,05	>0,05	>0,05	>0,05
	N	4	4	4	4	0

Improvement of astheno-neurotic syndrome in AIDS patients after Embryonic Stem Cell Transplantation

Feature	Whole group (N=38)		Patients with astheno-neurotic syndrome (N=22)
Feature	abs	%	abs	%
Increasing of energy level	17	45	14	64
Improving of appetites	13	34	11	50
Normalization of sleeping	15	39	13	59
Pain decrease	5	13	4	18
Diminishing of irritation	12	32	12	55
Increasing of work capability	7	18	6	27
Decreasing of fatigue	14	37	11	50
Improvement of mood	21	55	17	77

Changes of CD4+ and CD8+ lymphocytes in patients with acute decrease
of CD4+ (more that 100 CD4+ cells/ml within 6 months) prior to the Embryonic Stem Cell Transplantation(N=15)

	Period of observation before treatment (days)					Day after treatment		Period of observation after treatment (days)
	60	30	14	7	1	7	14	30	60
CD4+ M	205,7	219,8	164,1	130,9	136,8	147,4	188,7	206,2	197,5
m	44,6	40,7	48,0	44,6	40,4	34,0	35,7	56,7	82,2
p		>0,05	>0,05	<0,05		>0,05	>0,05	<0,05	<0,05
CD8+ M	829,7	977,5	1178	934,0	996,2	11560	1110	1088	873,0
m	221,9	279,9	314,8	258,6	237,3	313,6	301,7	323,1	350,3
p		>0,05	>0,05	>0,05		>0,05	>0,05	>0,05	>0,05

Changes of CD4+ lymphocytes in patients with acute decrease of CD4+ (more than 100 cellsml within 6 month) before and after the Transplantation of Embryonic Stem Cells (N=15)

Embryonic Stem Cells Transplantation combined with antiretroviral therapy

Some patients received antiretroviral therapy during several months, however without any results. As a result of development of resistance against AZT, these patients demonstrated negative changes of CD4+ lymphocytes count against the background of azido-timidin administration. Immunological monitoring in patients that received antiretroviral therapy prior to and after Embryonic Stem Cell Transplantation is shown below.

Changes of CD4+ lymphocytes in AIDS patients, that received antiretroviral therapy, prior to and after Embryonic Stem Cell Transplantation (N=10)

Stat. par.	Period of observation prior to Embryonic Stem Cells Transplantation (days)					Day after ESCT		Period of observation after Embryonic Stem Cells Transplantation (days)
Stat. par.	60	30	14	7	1	7	14	30	60
M	260,7	223,8	160,5	96,5	110,4	146,9	180,7	199,9	197,5
m	60,9	45,1	55,3	25,7	33,6	37,5	38,4	62,2	82,2
Min	41	44	30	8	9	41	41	47	22
Max	614	431	440	189	376	429	389	700	533
p						0,025	0,017	0,068	0,054

Changes of CD4+ lymphocytes in AIDS patients received antiretroviral therapy before and after the Transplantation of Embryonic Stem Cells (N=10)

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