Repeated RBD boosting leads to immune tolerance

And persistent infections "galore"

The following paper was published at the end of last year, and I missed it. For further context here are my observations on why using RBD as a vaccine target ain’t the best idea. As a precaution, everything I will discuss here in relation to the immune system is “fixable”, especially in regards to CD8 T Cells (the cells that kill the bad stuff inside you, simplified).

More "domain specific" vaccines and XBB

Moderna new "domain specific" vaccine

To boost or not to boost after infection ?

Extended SARS-CoV-2 RBD booster vaccination induces humoral and cellular immune tolerance in mice

Extended immunization did not enhance RBD specific antibody production in mice

However, subsequent immunization gradually reduced the titer of RBD-specific IgG antibodies, and a significant difference could be detected after the second injection of RBD booster vaccines

Because serum IgG subclass distribution is indicative of Th1- or Th2 (T helper cells, Th) biased immunity, we analyzed the IgG subclass antibody responses induced by the RBD vaccine. ELISA results showed that both RBD-specific IgG1 and IgG2a were detected in the serum from immunized mice. IgG1 titer was significantly higher than that of IgG2a, indicating that the RBD vaccine induced a Th2-like response by preferentially potentiating serum IgG1 antibody.

IgG1 titer in mice immunized by multiple boosters was significantly lower than that without booster. These results suggested that the RBD vaccine could stimulate the production of RBD-specific antibodies with a dominance of the Th2-type, whereas the addition of RBD booster vaccines did not enhance RBD specific antibody production in mice.

In line with much of the evidence covered so far, now we are presented with even more evidence on how foolish it would be to vaccinate with an RBD-based design, one that is becoming in vogue in recent new vaccine research tests. Boosting with an RBD-based vaccine after 4 doses (therefore making the RBD one the fifth booster) didn’t have the desired effect, in turn, the more you were vaccinated with it, the lower the response became. And since not all immune responses are created equal, they went further to categorize which type of immune response this vaccine generated.

Here is where it gets “dark”. The further boosting of the RBD vaccine induced a Th2 dominant response, and if you are not a long-time reader, a Th2 dominant state is by definition an “allergic” state, without actually increasing antibody production the more you “boost”. Why this is dark ?

Because this is precisely one of the “catastrophic” scenarios Ralph Baric, father of coronavirus research stated years ago in regards to coronavirus vaccines, and something observed in actual trials for vaccines years ago. A similar mechanism is also a potential explanation for the IgG class switch to IgG4, and the further we go into this paper, the more sense these atypical changes make.

Extended immunization inhibited the production of RBD-specific memory B cells

The results showed that RBD recombination proteins could promote significant increases in the productions of IL-4 and IL-5, whereas the serum levels of either IL-4 or IL-5 were comparatively lower with extended vaccination. The above information demonstrated that additional RBD booster vaccines might result in a loss of RBD-specific humoral immunity and promote the immune tolerance.

The production of specific cytokines is diminished upon excessive vaccination, and taken together, boosting too many results in loss of RBD-specific immunity and promoted immune tolerance. Immune tolerance means your body doesn’t respond too well (or at all) to antigens, the little lego pieces that activate the cells in your body. The same applied to antibodies, the further the researchers boosted with the same protein, the fewer antibodies the body produce, in a state of tolerance and anergy.

To further elucidate the effects of extended boosting in the immune response and antibody production, researchers went to investigate the formation of germinal centers, where B cells differentiate and induce further specific immune responses. The results of this test showed that further boosters don’t elevate the germinal center responses and might tilt the immune system from immune response to tolerance. If the following section was covered months ago, I would be smiling or laughing, but now I just feel sad for being “right”.

Extended immunization inhibited the activation of CD4+T cell immune responses

With the observed disadvantages in the humoral immunity caused by extended immunization, we moved on to determine whether there were any differences in the cellular immune responses to the two vaccine courses.
A detailed study of the T cell subsets in the CD4+T cells revealed that the proportion of effector memory T cells (Tem) and central memory T cells (Tcm) in CD4+T cells in the extended group was sharply decreased relative to that from the normal group, along with relatively increases in naive T cells (Tn) population. Specially, we observed that the frequency of CD4+ Te cells significant elevates in the extended group.

The results from flow cytometric analysis showed that a higher percentage of CD4+ CD25+ foxp-3+ Treg cells were detected in the extended vaccination group, compared with the normal group and the PBS control group

Further, we tested the serum level of IL-10, which was mainly secreted by Treg cells. The ELISA results showed that higher amount of IL-10 was detected in samples from the extended vaccination group than the other two groups, which was consistent with the increased percentile of Treg cells. These data suggested Treg cells might play an important role in the immune tolerances to the extended immunization with RBD vaccines.

Central to what I used to refer to as the “reverse AIDS hypothesis”, later changing the name to PAID (Paradoxical Acquired Immune Dysfunction, also a slight joke on the corruption behind it all) because of the convergence of the certain circles of “influencer” on pushing misleading narrative was the CD4/CD8 responses in relation to the mRNA vaccines.

The whole basis for effectively reverse engineering the mRNA vaccines to such a detailed point I got messages from insiders, and here we are presented with further evidence of a similar dynamic. While I research and read an abnormally large amount of science, I abstain from writing about certain topics for multiple reasons, not only the aforementioned influencer one. The IL-10/T-regs are one of them.

This specific, persistent, lasting immune state is often seen in a small subset of both recovered severe Covid patients and in quite a few numbers of Long Covid, among other pathologies. Since the dominant immune response towards both SARS-CoV-2, the Spike Protein and mRNA are effectively the same, a Th17 one (highly inflammatory), you also don’t want excessive IL-10, high expression of T-Regs, this will also create long-term (autoimmune, chronic infection) problems. The following section would make me smile even more months back.

Extended immunization inhibited CD8+ T cell-mediated immune response

It has been reported that repeated antigen stimulation induces the exhaustion of CD8+T cells; therefore, we tested whether there were any differences in exhaustion marker levels between two immunization courses. We found that the cell surface expressions of PD-1 and LAG-3 on CD8+T cells from mouse splenocytes were evidently higher in the extended vaccination group, comparing to either the conventional group or the PBS control. Concomitantly, the proportion of PD-1- LAG-3- CD8+T cells in the extended group was significantly less than the other groups . The expressions of PD-1 and LAG-3 on the Te subsets of CD8+T cells were further analyzed, and we found that the highest level of LAG-3 was expressed in the Te subsets of CD8+T cells from samples with prolonged immunization . These data indicated that continues administration of RBD booster vaccines could lead to reduced CD8+T cell activation with increased exhaustion. Overall, our findings evidenced the potential risk of adaptive immune tolerance from prolonged course of immunization with homologous vaccine boosters, and suggested that the applications of multiple booster vaccines with protective intent should be preceded with caution.

This section is a statement to many substacks I wrote while refining our hypothesis (mine and from a friend, of which I always give credit). There is a large body of evidence not merely related to SARS-CoV-2 but to many other types of infections, both acute or chronic demonstrating how critical a CD8+ response is, this is how your body deals and clean itself from any type of pathogen among other things.

A lack of proper CD8 response towards anything will lead to improper immune response, and your body won’t be able to clean the infection properly, which the following papers after this one will serve as examples of why and how this is a terrible outcome, the further we go. The discussion section makes some pertinent arguments.

Together with a diminished CD8+ response, improper production of IL-2, but especially Interferon-Gamma (IFN-y) an assured way to not only be aware of latent infections, but create persistent, long, and exacerbate chronic infections from a myriad of pathogens. This is a point the authors make further down in this paragraph, over vaccination can severely impact the immune responses and drive disease severity of the ones getting reinfected, which at this point is basically most of the planet.

I will further explain why these dynamics are so bad from many points, but in regards to Long Covid and other complex dynamics, it can be really bad, but also for the “average person”. Being tolerant to the virus doesn’t mean your body won’t “touch” it, it means your body will interact with it but won’t have a proper response, which is one of the three basis to create an autoimmune state in regards to SARS-CoV-2. See any of my substacks (including the recent one about the N protein and MS) to understand why this improper response can be significantly worse the further down the line we go.

Before going further I will leave the reader with this paragraph from the authors.

Although RBD subunit vaccines cannot entirely represent inactivated or mRNA vaccines, especially in antigen delivery way. A recent report in The New England Journal of Medicine demonstrated that a fourth mRNA vaccination of healthy young health care workers only shows marginal benefits (Regev-Yochay et al., 2022). Whether extended vaccination with other COVID-19 vaccines based on wild-type SARS-CoV-2 sequence will induce immune tolerance, further investigations are required.

What we just covered here will also most likely lead to this.

Persistent SARS-CoV-2 infection in patients seemingly recovered from COVID-19

SARS-CoV-2 infection is clinically heterogeneous, ranging from asymptomatic to deadly. A few patients with COVID-19 appear to recover from acute viral infection but nevertheless progress in their disease and eventually die, despite persistent negativity at molecular tests for SARS-CoV-2 RNA. Here, we performed post-mortem analyses in 27 consecutive patients who had apparently recovered from COVID-19 but had progressively worsened in their clinical conditions despite repeated viral negativity in nasopharyngeal swabs or bronchioalveolar lavage for 11–300 consecutive days (average: 105.5 days). Three of these patients remained PCR-negative for over 9 months. Post-mortem analysis revealed evidence of diffuse or focal interstitial pneumonia in 23/27 (81%) patients, accompanied by extensive fibrotic substitution in 13 cases (47%). Despite apparent virological remission, lung pathology was similar to that observed in acute COVID-19 individuals, including micro- and macro-vascular thrombosis (67% of cases), vasculitis (24%), squamous metaplasia of the respiratory epithelium (30%), frequent cytological abnormalities and syncytia (67%), and the presence of dysmorphic features in the bronchial cartilage (44%). Consistent with molecular test negativity, SARS-CoV-2 antigens were not detected in the respiratory epithelium. In contrast, antibodies against both spike and nucleocapsid revealed the frequent (70%) infection of bronchial cartilage chondrocytes and para-bronchial gland epithelial cells. In a few patients (19%), we also detected positivity in vascular pericytes and endothelial cells. Quantitative RT-PCR amplification in tissue lysates confirmed the presence of viral RNA. Together, these findings indicate that SARS-CoV-2 infection can persist significantly longer than suggested by standard PCR-negative tests, with specific infection of specific cell types in the lung. Whether these persistently infected cells also play a pathogenic role in long COVID remains to be addressed

I personally saw many of my family members with persistent lung “symptoms” including my own mother for weeks after mild Covid infections and could observe all around me in public, and I know many of my Twitter followers and others observed the same. What we discussed above is an assured way to create what is described in this paper, and this isn’t exactly new and is an attempt to demonstrate how complex many of these pathways are, here is a piece covering specifically persistent infected people.

More antigenic sin, immune shift in persistent infection

Very early on I already suspect persistent infection was happening at a larger scale than thought by most people by just analyzing the pathways the mRNA induces and messes with, so most of my supplements suggestion are, in a covert way, “made” to bring balance to a skewed immune system and properly clean any sort of infection. Of course, each person is unique and blanket suggestions for large amounts of people only go so far, but for the most part, the majority of people had a pretty good response and recovery.

When dealing with the states described in this substack one should pay more attention to both antioxidant intakes (consumption) and special attention to mitochondrial health, because at the core of all these different dysfunctional states lies a poor energy generation/efficiency conundrum. Dealing with this cascade positively into dealing with said states and kick start the immune system into proper function.

On one side I am not happy about being right, on the other side I am because at least not only I was on the right path, but most of what I research to try and help people is effective in doing so.


As always, I am really grateful to anyone who chooses to support my substack or when they decided to use Kofi and anyone who shares what they find useful.